Il-13 binding protein

ABSTRACT

The present invention relates to antigen binding proteins to human IL-13, including anti-IL-13 antibodies and anti-IL-3/anti-IL-4 mAbdAbs, pharmaceutical formulations containing them and to the use of such antigen binding proteins in the treatment and/or prophylaxis of inflammatory diseases such as asthma or IPF.

FIELD OF THE INVENTION

The present invention relates to antigen binding proteins, particularlyantibodies that bind to interleukin 13 (IL-13) and neutralise theactivity thereof, polynucleotides encoding such antigen bindingproteins, pharmaceutical formulations containing said antigen bindingproteins and to the use of such antigen binding proteins in thetreatment and/or prophylaxis of diseases associated with inflammation,such as asthma. Other aspects, objects and advantages of the presentinvention will become apparent from the description below.

BACKGROUND OF THE INVENTION Interleukin-13 (IL-13)

IL-13 is a 12 kDa secreted cytokine originally described as a Tcell-derived cytokine that inhibits inflammatory cytokine production.Structural studies indicate that it has a four-helical bundlearrangement held by two disulphide bonds. Although IL-13 has fourpotential glycosylation sites, analysis of native IL-13 from rat lunghas indicated that it is produced as an unglycosylated molecule.Expression of human IL-13 from NSO and COS-7 cells confirms thisobservation (Eisenmesser et al, J. Mol. Biol. 2001 310(1):231-241; Moyet al, J. Mol. Biol. 2001 310(1):219-230; Cannon-Carlson et al, ProteinExpression and Purification 1998 12(2):239-248).

IL-13 has been implicated in asthma, Chronic Obstructive PulmonaryDisease (COPD), Allergic disease including atopic dermatitis andallergic rhinitis, Esophagal eosinophilia, Oncology Indications, e.g.B-cell chronic lymphocytic leukemia (B-CLL) and Hodgkin's disease,Inflammatory Bowel Diseases e.g. ulcerative colitis, Crohn's disease andindeterminate colitis, Psoriasis and Psoriatic Arthritis, Acutegraft-versus-host disease, Diabetic nephropathy, Fibrotic Conditionssuch as Pulmonary fibrosis e.g. Idiopathic Pulmonary Fibrosis (IPF).

SUMMARY OF INVENTION

The invention provides antigen binding proteins which bind to IL-13, forexample IL-13 antibodies, and to the combination of such IL-13antibodies with an IL-4 antagonist and/or an IL-5 antagonist. The IL-13antibodies of the present invention are related to, or derived from, amurine mAb 6A1, wherein the CDRH3 is mutated. The 6A1 murine heavy chainvariable region amino acid sequence is provided as SEQ ID NO: 58. The6A1 murine light chain variable region amino acid sequence is providedas SEQ ID NO 59.

The heavy chain variable regions (VH) of the present invention comprisethe following CDRs (as defined by Kabat (Kabat et al; Sequences ofproteins of Immunological Interest NIH, 1987)):

The CDRs of the heavy chain variable regions of the present inventionmay comprise the following CDRs:

CDR According to Kabat H1 DTYMH (SEQ ID NO: 1) H2 TIDPANGNTKYVPKFQG (SEQID NO: 2) H3 WIYDDYHYDDYYAMDY (SEQ ID NO: 4); or SVYDDYHYDDYYAMDY (SEQID NO: 5); or SIFDDYHYDDYYAMDY (SEQ ID NO: 6); or SIYEDYHYDDYYAMDY (SEQID NO: 7); or SIYDDYAYDDYYAMDY (SEQ ID NO: 8); or SIYDDYEYDDYYAMDY (SEQID NO: 9); or SIYDDYQYDDYYAMDY (SEQ ID NO: 10); or SIYDDYRYDDYYAMDY (SEQID NO: 11); or SIYDDYSYDDYYAMDY (SEQ ID NO: 12); or SIYDDYTYDDYYAMDY(SEQ ID NO: 13) or SIYDDYVYDDYYAMDY (SEQ ID NO: 14); or SIYDDYHADDYYAMDY(SEQ ID NO: 15); or SIYDDYHIDDYYAMDY (SEQ ID NO: 16); orSIYDDYHWDDYYAMDY (SEQ ID NO: 17); or SIYDDYHVDDYYAMDY (SEQ ID NO: 18)

The light chain variable regions of the present invention comprise thefollowing CDRs (as defined by Kabat):

CDR According to Kabat L1 RSSQNIVHINGNTYLE (SEQ ID NO: 19) L2 KISDRFS(SEQ ID NO: 20) L3 FQGSHVPWT (SEQ ID NO: 21)

The CDR sequences of antibodies can be determined by the Kabat numberingsystem (Kabat et al; Sequences of proteins of Immunological InterestNIH, 1987), as set out in the tables above, alternatively they can bedetermined using the Chothia numbering system (Al-Lazikani et al.,(1997) JMB 273, 927-948), the contact definition method (MacCallum R.M., and Martin A. C. R. and Thornton J. M, (1996), Journal of MolecularBiology, 262 (5), 732-745) or any other established method for numberingthe residues in an antibody and determining CDRs known to the skilledman in the art. The CDRs of the invention described herein may bedefined by any of these methods, or by using a combination of Chothiaand Kabat numbering, for example CDRH1 may be defined as comprisingFYIKDTYMH (SEQ ID NO 60) or GFYIKDTYMH (SEQ ID NO 61).

The present invention also provides an antigen-binding proteincomprising the IL-13 antibody of the present invention which is linkedto one or more epitope-binding domains, for example an antigen-bindingprotein comprising the IL-13 antibody of the present invention linked toan epitope-binding domain which is capable of binding to IL-4, or anantigen-binding protein comprising the IL-13 antibody of the presentinvention linked to an epitope-binding domain which is capable ofbinding to IL-5, or an antigen-binding protein comprising the IL-13antibody of the present invention linked to a first epitope-bindingdomain which is capable of binding to IL-4 and a second epitope-bindingdomain which is capable of binding to IL-5.

The present invention also provides a method of decreasing theaggregation propensity of an immunoglobulin single variable domain, forexample a human dAb, for example human VK domain antibody, by mutationof residue 89 (kabat numbering) to a residue selected from ‘Q’(glutamine), ‘G’ (glycine), ‘S’ (serine), ‘M’ (methionine), ‘A’(alanine), ‘T’ (threonine) and ‘E’ (glutamic acid). In one embodiment,the method involves mutation of residue 89 (kabat numbering) to aresidue selected from ‘Q’ (glutamine), ‘G’ (glycine), ‘S’ (serine), ‘M’(methionine) and ‘E’ (glutamic acid). In a further embodiment the methodinvolves mutation of residue 89 (kabat numbering) to ‘Q’ (glutamine).

In one embodiment this method can be applied to the anti-IL-4 domainantibody of SEQ ID NO: 80, resulting in a mutated dAb sequence, forexample SEQ ID NO:94. Such mutated dAbs may be alone or as part of alarger sequence, for example part of a mAbdAb sequence, resulting in forexample, a dAb comprising a sequence selected from SEQ ID NO: 117-134.

The invention also provides human VK dAbs which have improvedaggregation profiles, for example a human VK dAb derived from a germlineframework selected from IGKV1-17, IGKV1D-17, IGKV1/OR2-108, IGKV1-6,IGKV5-2, IGKV1D-42, IGKV2-24, IGKV2-28, IGKV2-30, IGKV2-40, IGKV2D-29,IGKV2D-30, IGKV2D-24 and IGKV6-21 wherein residue 89 (kabat numbering)of the VK dAb is ‘Q’ (glutamine). In one such embodiment the VK dAbcomprises germline framework regions selected from the germlineframeworks of IGKV1-17, IGKV1D-17, IGKV1/OR2-108, IGKV1-6, IGKV5-2,IGKV1D-42, IGKV2-24, IGKV2-28, IGKV2-30, IGKV2-40, IGKV2D-29, IGKV2D-30,IGKV2D-24 and IGKV6-21 wherein residue 89 (kabat numbering) of the VKdAb is ‘Q’ (glutamine).

In one embodiment, the invention provides a human dAb comprising thesequence of SEQ ID NO: 80 which comprises a mutation at position 89(kabat numbering) wherein the mutated position 89 is selected from ‘Q’(glutamine), ‘G’ (glycine), ‘S’ (serine), ‘M’ (methionine), ‘A’(alanine), ‘T’ (threonine) and ‘E’ (glutamic acid). For example theinvention provides a human dab comprising the sequence of SEQ ID NO:80wherein position 89 (kabat numbering) is mutated to a residue selectedfrom ‘Q’ (glutamine), ‘G’ (glycine), ‘S’ (serine), ‘M’ (methionine) and‘E’ (glutamic acid).

In one embodiment the invention provides a human dAb comprising thesequence of SEQ ID NO: 94

The invention also provides a polynucleotide sequence encoding a heavychain of any of the antigen-binding proteins described herein, and apolynucleotide encoding a light chain of any of the antigen-bindingproteins described herein. Such polynucleotides represent the codingsequence which corresponds to the equivalent polypeptide sequences,however it will be understood that such polynucleotide sequences couldbe cloned into an expression vector along with a start codon, anappropriate signal sequence and a stop codon.

The invention also provides a recombinant transformed or transfectedhost cell comprising one or more polynucleotides encoding a heavy chainand a light chain of any of the antigen-binding proteins describedherein.

The invention further provides a method for the production of any of theantigen-binding proteins described herein which method comprises thestep of culturing a host cell comprising a first and second vector, saidfirst vector comprising a polynucleotide encoding a heavy chain of anyof the antigen-binding proteins described herein and said second vectorcomprising a polynucleotide encoding a light chain of any of theantigen-binding proteins described herein, in a suitable culture media,for example serum-free culture media.

The invention further provides a pharmaceutical composition comprisingan antigen-binding protein as described herein a pharmaceuticallyacceptable carrier.

In a further aspect, the present invention provides a method oftreatment or prophylaxis of diseases or disorders associated with atopicdiseases/disorders and chronic inflammatory diseases/disorders byadministration of the antigen binding protein of the present invention.Of particular interest is their use in the treatment of asthma, such asallergic asthma, particularly severe asthma (that is asthma that isunresponsive to current treatment, including systemically administeredcorticosteroids; see Busse W W et al, J. Allergy Clin. Immunol 2000,106: 1033-1042), “difficult” asthma (defined as the asthmatic phenotypecharacterised by failure to achieve control despite maximallyrecommended doses of prescribed inhaled steroids, see Barnes P J (1998),Eur Respir J 12:1208-1218), “brittle” asthma (defines a subgroup ofpatients with severe, unstable asthma who maintain a wide peakexpiratory flow (PEF) variability despite high doses of inhaledsteroids, see Ayres J G et al (1998) Thorax 58:315-321), nocturnalasthma, premenstrual asthma, steroid resistant asthma (see Woodcock A J(1993) Eur Respir J 6:743-747), steroid dependent asthma (defined asasthma that can be controlled only with high doses of oral steroids),aspirin induced asthma, adult-onset asthma, paediatric asthma.Antibodies of the invention maybe used to prevent, reduce the frequencyof, or mitigate the effects of acute, asthmatic episodes (statusasthmaticus). Antibodies of the invention may also be used to reduce thedosing required (either in terms of amount administered or frequency ofdosing) of other medicaments used in the treatment of asthma. Forexample, antibodies of the invention may be used to reduce the dosingrequired for steroid treatment of asthma such as corticosteroidtreatment (“steroid sparing”). Other diseases or disorders that may betreated with antibodies of the invention include atopic dermatitis,allergic rhinitis, Crohn's disease, chronic obstructive pulmonarydisease (COPD), eosinophilic esophagitis, fibrotic diseases or disorderssuch as idiopathic pulmonary fibrosis, progressive systemic sclerosis(scleroderma), hepatic fibrosis, hepatic granulomas, schistosomiasis,leishmaniasis, and diseases of cell cycle regulation, e.g. Hodgkinsdisease, B cell chronic lymphocytic leukaemia.

In another aspect, the invention provides the use of an antigen bindingprotein of the invention in the preparation of a medicament fortreatment or prophylaxis of atopic diseases/disorders and chronicinflammatory diseases/disorders. Of particular interest is their use inthe treatment of asthma, such as allergic asthma, particularly severeasthma (that is asthma that is unresponsive to current treatment,including systemically administered corticosteroids; see Busse W W etal, J. Allergy Clin. Immunol 2000, 106: 1033-1042), “difficult” asthma(defined as the asthmatic phenotype characterised by failure to achievecontrol despite maximally recommended doses of prescribed inhaledsteroids, see Barnes P J (1998), Eur Respir J 12:1208-1218), “brittle”asthma (defines a subgroup of patients with severe, unstable asthma whomaintain a wide peak expiratory flow (PEF) variability despite highdoses of inhaled steroids, see Ayres J G et al (1998) Thorax58:315-321), nocturnal asthma, premenstrual asthma, steroid resistantasthma (see Woodcock A J (1993) Eur Respir J 6:743-747), steroiddependent asthma (defined as asthma that can be controlled only withhigh doses of oral steroids), aspirin induced asthma, adult-onsetasthma, paediatric asthma. Antibodies of the invention maybe used toprevent, reduce the frequency of, or mitigate the effects of acute,asthmatic episodes (status asthmaticus). Antibodies of the invention mayalso be used to reduce the dosing required (either in terms of amountadministered or frequency of dosing) of other medicaments used in thetreatment of asthma. For example, antibodies of the invention may beused to reduce the dosing required for steroid treatment of asthma suchas corticosteroid treatment (“steroid sparing”). Other diseases ordisorders that may be treated with antibodies of the invention includeatopic dermatitis, allergic rhinitis, Crohn's disease, chronicobstructive pulmonary disease (COPD), eosinophilic esophagitis, fibroticdiseases or disorders such as idiopathic pulmonary fibrosis, progressivesystemic sclerosis (scleroderma), hepatic fibrosis, hepatic granulomas,schistosomiasis, leishmaniasis, and diseases of cell cycle regulation,e.g. Hodgkins disease, B cell chronic lymphocytic leukaemia.

Other aspects and advantages of the present invention are describedfurther in the detailed description and the embodiments thereof.

DEFINITIONS

The term “binds to human IL-13” as used throughout the presentspecification in relation to antigen binding proteins thereof of theinvention means that the antigen binding protein binds human IL-13(hereinafter referred to as hIL-13) with no or insignificant binding toother human proteins such as IL-4. In particular the antigen bindingproteins of the present invention bind to human IL-13 in that they canbe seen to bind to human IL-13 in a Biacore assay (for example theBiacore assay described in example 3). The term however does not excludethe fact that certain antigen binding proteins of the invention may alsobe cross-reactive with IL-13 from other species, for example cynomolgusIL-13.

The term “antigen binding protein” as used herein refers to antibodies,antibody fragments and other protein constructs which are capable ofbinding to and neutralising human IL-13.

The terms Fv, Fc, Fd, Fab, or F(ab)₂ are used with their standardmeanings (see, e.g., Harlow et al., Antibodies A Laboratory Manual, ColdSpring Harbor Laboratory, (1988)).

A “chimeric antibody” refers to a type of engineered antibody whichcontains a naturally-occurring variable region (light chain and heavychains) derived from a donor antibody in association with light andheavy chain constant regions derived from an acceptor antibody.

A “humanised antibody” refers to a type of engineered antibody havingits CDRs derived from a non-human donor immunoglobulin, the remainingimmunoglobulin-derived parts of the molecule being derived from one (ormore) human immunoglobulin(s). In addition, framework support residuesmay be altered to preserve binding affinity (see, e.g., Queen et al.,Proc. Natl. Acad Sci USA, 86:10029-10032 (1989), Hodgson et al.,Bio/Technology, 9:421 (1991)). A suitable human acceptor antibody may beone selected from a conventional database, e.g., the KABAT® database,Los Alamos database, and Swiss Protein database, by homology to thenucleotide and amino acid sequences of the donor antibody. A humanantibody characterized by a homology to the framework regions of thedonor antibody (on an amino acid basis) may be suitable to provide aheavy chain constant region and/or a heavy chain variable frameworkregion for insertion of the donor CDRs. A suitable acceptor antibodycapable of donating light chain constant or variable framework regionsmay be selected in a similar manner. It should be noted that theacceptor antibody heavy and light chains are not required to originatefrom the same acceptor antibody. The prior art describes several ways ofproducing such humanised antibodies—see for example EP-A-0239400 andEP-A-054951

The term “donor antibody” refers to an antibody (monoclonal, and/orrecombinant) which contributes the amino acid sequences of its variableregions, CDRs, or other functional fragments or analogs thereof to afirst immunoglobulin partner, so as to provide the alteredimmunoglobulin coding region and resulting expressed altered antibodywith the antigenic specificity and neutralizing activity characteristicof the donor antibody.

The term “acceptor antibody” refers to an antibody (monoclonal and/orrecombinant) heterologous to the donor antibody, which contributes all(or any portion, but in some embodiments all) of the amino acidsequences encoding its heavy and/or light chain framework regions and/orits heavy and/or light chain constant regions to the firstimmunoglobulin partner. In certain embodiments a human antibody is theacceptor antibody.

“CDRs” are defined as the complementarity determining region amino acidsequences of an antibody which are the hypervariable regions ofimmunoglobulin heavy and light chains. See, e.g., Kabat et al.,Sequences of Proteins of Immunological Interest, 4th Ed., U.S.Department of Health and Human Services, National Institutes of Health(1987). There are three heavy chain and three light chain CDRs (or CDRregions) in the variable portion of an immunoglobulin. Thus, “CDRs” asused herein refers to all three heavy chain CDRs, or all three lightchain CDRs (or both all heavy and all light chain CDRs, if appropriate).The structure and protein folding of the antibody may mean that otherresidues are considered part of the antigen binding region and would beunderstood to be so by a skilled person. See for example Chothia et al.,(1989) Conformations of immunoglobulin hypervariable regions; Nature342, p877-883.

As used herein the term “domain” refers to a folded protein structurewhich has tertiary structure independent of the rest of the protein.Generally, domains are responsible for discrete functional properties ofproteins and in many cases may be added, removed or transferred to otherproteins without loss of function of the remainder of the protein and/orof the domain. An “antibody single variable domain” is a foldedpolypeptide domain comprising sequences characteristic of antibodyvariable domains. It therefore includes complete antibody variabledomains and modified variable domains, for example, in which one or moreloops have been replaced by sequences which are not characteristic ofantibody variable domains, or antibody variable domains which have beentruncated or comprise N- or C-terminal extensions, as well as foldedfragments of variable domains which retain at least the binding activityand specificity of the full-length domain.

The phrase “immunoglobulin single variable domain” refers to an antibodyvariable domain (V_(H), V_(HH), V_(L)) that specifically binds anantigen or epitope independently of a different V region or domain. Animmunoglobulin single variable domain can be present in a format (e.g.,homo- or hetero-multimer) with other, different variable regions orvariable domains where the other regions or domains are not required forantigen binding by the single immunoglobulin variable domain (i.e.,where the immunoglobulin single variable domain binds antigenindependently of the additional variable domains). A “domain antibody”or “dAb” is the same as an “immunoglobulin single variable domain” whichis capable of binding to an antigen as the term is used herein. Animmunoglobulin single variable domain may be a human antibody variabledomain, but also includes single antibody variable domains from otherspecies such as rodent (for example, as disclosed in WO 00/29004), nurseshark and Camelid V_(HH) dAbs (nanobodies). Camelid V_(HH) areimmunoglobulin single variable domain polypeptides that are derived fromspecies including camel, llama, alpaca, dromedary, and guanaco, whichproduce heavy chain antibodies naturally devoid of light chains. SuchV_(HH) domains may be humanised according to standard techniquesavailable in the art, and such domains are still considered to be“domain antibodies” according to the invention. As used herein “V_(H)includes camelid V_(HH) domains. NARV are another type of immunoglobulinsingle variable domain which were identified in cartilaginous fishincluding the nurse shark. These domains are also known as Novel AntigenReceptor variable region (commonly abbreviated to V(NAR) or NARV). Forfurther details see Mol. Immunol. 44, 656-665 (2006) and US20050043519A.

The term “Epitope-binding domain” refers to a domain that specificallybinds an antigen or epitope independently of a different V region ordomain, this may be a domain antibody (dAb), for example a human,camelid or shark immunoglobulin single variable domain or it may be adomain which is a derivative of a non-Immunoglobulin scaffold, forexample a non-immunoglobulin scaffold selected from the group consistingof CTLA-4 (Evibody); lipocalin; Protein A derived molecules such asZ-domain of Protein A (Affibody, SpA), A-domain (Avimer/Maxibody); Heatshock proteins such as GroEI and GroES; transferrin (trans-body);ankyrin repeat protein (DARPin); peptide aptamer; C-type lectin domain(Tetranectin); human γ-crystallin and human ubiquitin (affilins); PDZdomains; scorpion toxinkunitz type domains of human protease inhibitors;and fibronectin (adnectin); which has been subjected to proteinengineering in order to obtain binding to a ligand other than thenatural ligand.

CTLA-4 (Cytotoxic T Lymphocyte-associated Antigen 4) is a CD28-familyreceptor expressed on mainly CD4+ T-cells. Its extracellular domain hasa variable domain-like Ig fold. Loops corresponding to CDRs ofantibodies can be substituted with heterologous sequence to conferdifferent binding properties. CTLA-4 molecules engineered to havedifferent binding specificities are also known as Evibodies. For furtherdetails see Journal of Immunological Methods 248 (1-2), 31-45 (2001)

Lipocalins are a family of extracellular proteins which transport smallhydrophobic molecules such as steroids, bilins, retinoids and lipids.They have a rigid 13-sheet secondary structure with a numer of loops atthe open end of the conical structure which can be engineered to bind todifferent target antigens. Anticalins are between 160-180 amino acids insize, and are derived from lipocalins. For further details see BiochimBiophys Acta 1482: 337-350 (2000), U.S. Pat. No. 7,250,297B1 andUS20070224633

An affibody is a scaffold derived from Protein A of Staphylococcusaureus which can be engineered to bind to antigen. The domain consistsof a three-helical bundle of approximately 58 amino acids. Librarieshave been generated by randomisation of surface residues. For furtherdetails see Protein Eng. Des. Sel. 17, 455-462 (2004) and EP1641818A1

Avimers are multidomain proteins derived from the A-domain scaffoldfamily. The native domains of approximately 35 amino acids adopt adefined disulphide bonded structure. Diversity is generated by shufflingof the natural variation exhibited by the family of A-domains. Forfurther details see Nature Biotechnology 23(12), 1556-1561 (2005) andExpert Opinion on Investigational Drugs 16(6), 909-917 (June 2007)

A transferrin is a monomeric serum transport glycoprotein. Transferrinscan be engineered to bind different target antigens by insertion ofpeptide sequences in a permissive surface loop. Examples of engineeredtransferrin scaffolds include the Trans-body. For further details see J.Biol. Chem. 274, 24066-24073 (1999).

Designed Ankyrin Repeat Proteins (DARPins) are derived from Ankyrinwhich is a family of proteins that mediate attachment of integralmembrane proteins to the cytoskeleton. A single ankyrin repeat is a 33residue motif consisting of two α-helices and a β-turn. They can beengineered to bind different target antigens by randomising residues inthe first α-helix and a β-turn of each repeat. Their binding interfacecan be increased by increasing the number of modules (a method ofaffinity maturation). For further details see J. Mol. Biol. 332, 489-503(2003), PNAS100(4), 1700-1705 (2003) and J. Mol. Biol. 369, 1015-1028(2007) and US20040132028A1.

Fibronectin is a scaffold which can be engineered to bind to antigen.Adnectins consists of a backbone of the natural amino acid sequence ofthe 10th domain of the 15 repeating units of human fibronectin type III(FN3). Three loops at one end of the β-sandwich can be engineered toenable an Adnectin to specifically recognize a therapeutic target ofinterest. For further details see Protein Eng. Des. Sel. 18, 435-444(2005), US20080139791, WO2005056764 and U.S. Pat. No. 6,818,418B1.

Peptide aptamers are combinatorial recognition molecules that consist ofa constant scaffold protein, typically thioredoxin (TrxA) which containsa constrained variable peptide loop inserted at the active site. Forfurther details see Expert Opin. Biol. Ther. 5, 783-797 (2005).

Microbodies are derived from naturally occurring microproteins of 25-50amino acids in length which contain 3-4 cysteine bridges—examples ofmicroproteins include KalataB1 and conotoxin and knottins. Themicroproteins have a loop which can be engineered to include upto 25amino acids without affecting the overall fold of the microprotein. Forfurther details of engineered knottin domains, see WO2008098796.

Other epitope binding domains include proteins which have been used as ascaffold to engineer different target antigen binding properties includehuman γ-crystallin and human ubiquitin (affilins), kunitz type domainsof human protease inhibitors, PDZ-domains of the Ras-binding proteinAF-6, scorpion toxins (charybdotoxin), C-type lectin domain(tetranectins) are reviewed in Chapter 7—Non-Antibody Scaffolds fromHandbook of Therapeutic Antibodies (2007, edited by Stefan Dubel) andProtein Science 15:14-27 (2006). Epitope binding domains of the presentinvention could be derived from any of these alternative proteindomains.

As used herein, the term “antigen-binding site” refers to a site on aprotein which is capable of specifically binding to antigen, this may bea single domain, for example an epitope-binding domain, or it may bepaired VH/VL domains as can be found on a standard antibody. In someaspects of the invention single-chain Fv (ScFv) domains can provideantigen-binding sites.

The terms “mAbdAb” and dAbmAb” are used herein to refer toantigen-binding proteins of the present invention. The two terms can beused interchangeably, and are intended to have the same meaning as usedherein.

The term “antigen binding protein” as used herein refers to antibodies,antibody fragments, for example a domain antibody (dAb), ScFv, FAb,FAb₂, and other protein constructs which are capable of binding toIL-13. Antigen binding molecules may comprise at least one Ig variabledomain, for example antibodies, domain antibodies, Fab, Fab′, F(ab′)2,Fv, ScFv, diabodies, mAbdAbs, affibodies, heteroconjugate antibodies orbispecifics. In one embodiment the antigen binding molecule is anantibody. In another embodiment the antigen binding molecule is a dAb,i.e. an immunoglobulin single variable domain such as a VH, VHH or VLthat specifically binds an antigen or epitope independently of adifferent V region or domain. Antigen binding molecules may be capableof binding to two targets, I.e. they may be dual targeting proteins.Antigen binding molecules may be a combination of antibodies and antigenbinding fragments such as for example, one or more domain antibodiesand/or one or more ScFvs linked to a monoclonal antibody. Antigenbinding molecules may also comprise a non-Ig domain for example a domainwhich is a derivative of a scaffold selected from the group consistingof CTLA-4 (Evibody); lipocalin; Protein A derived molecules such asZ-domain of Protein A (Affibody, SpA), A-domain (Avimer/Maxibody); Heatshock proteins such as GroEI and GroES; transferrin (trans-body);ankyrin repeat protein (DARPin); peptide aptamer; C-type lectin domain(Tetranectin); human γ-crystallin and human ubiquitin (affilins); PDZdomains; scorpion toxinkunitz type domains of human protease inhibitors;and fibronectin (adnectin); which has been subjected to proteinengineering in order to obtain binding to IL-13. As used herein “antigenbinding protein” will be capable of antagonising and/or neutralisinghuman IL-13. In addition, an antigen binding protein may block IL-13activity by binding to IL-13 and preventing a natural ligand frombinding and/or activating the receptor.

As used herein “IL-13 antagonist” includes any compound capable ofreducing and or eliminating at least one activity of IL-13. By way ofexample, an IL-13 antagonist may bind to IL-13 and that binding maydirectly reduce or eliminate IL-13 activity or it may work indirectly byblocking at least one ligand from binding the receptor.

As used herein “IL-4 antagonist” includes any compound capable ofreducing and or eliminating at least one activity of IL-4. By way ofexample, an IL-4 antagonist may bind to IL-4 and that binding maydirectly reduce or eliminate IL-4 activity or it may work indirectly byblocking at least one ligand from binding the receptor.

As used herein “IL-5 antagonist” includes any compound capable ofreducing and or eliminating at least one activity of IL-5. By way ofexample, an IL-5 antagonist may bind to IL-5 and that binding maydirectly reduce or eliminate IL-5 activity or it may work indirectly byblocking at least one ligand from binding the receptor.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment the antigen binding proteins of the present inventioncomprise a heavy chain variable region containing a CDRH3 selected fromthe list consisting of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ IDNO: 7, SEQ ID NO:8 and SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQID NO: 12, SEQ ID NO: 13, SEQ ID NO:14 and SEQ ID NO: 15, SEQ ID NO: 16,SEQ ID NO: 17 and SEQ ID NO: 18 and a suitable CDRH1 and CDRH2, pairedwith a light chain variable region containing a suitable CDRL1, CDRL2and CDRL3 to form an antigen binding Fv unit which binds to human IL-13.In one embodiment the antigen binding proteins of the present inventionneutralise the activity of human IL-13

In one aspect of this embodiment the CDRH1 as set out in SEQ ID NO: 1 orSEQ ID NO: 60, or SEQ ID NO:61 and CDRH2 as set out in SEQ ID NO: 2 arealso present in the heavy chain variable region. In a further aspect ofthis embodiment the CDRHL1 as set out in SEQ ID NO: 19, CDRL2 as set outin SEQ ID NO:20 and CDRL3 as set out in SEQ ID NO: 21 are also presentin the light chain variable region.

In another aspect the antigen binding protein binds to human IL-13 withhigh affinity as measured by Biacore of 10 nM or less, and moreparticularly 2 nM or less, for example between about 0.8 nM and 2 nM, 1nM or less, or 100 pM or less, for example between about 20 pM and about100 pM or between about 20 pM and about 80 pM, or between about 20 pMand about 60 pM. In one such embodiment, this is measured by Biacorewith the antigen binding protein being captured on the biosensor chip,for example as set out in Example 3.

The heavy chain variable regions of the present invention may beformatted together with light chain variable regions to allow binding tohuman IL-13, in the conventional immunoglobulin manner (for example,human IgG, IgA, IgM etc.) or in any other “antibody-like” format thatbinds to human IL-13 (for example, single chain Fv, diabodies, Tandabs™etc (for a summary of alternative “antibody” formats see Holliger andHudson, Nature Biotechnology, 2005, Vol 23, No. 9, 1126-1136)).

The antigen binding proteins of the present invention are derived fromthe murine antibody having the variable regions as described in SEQ IDNO:58 and SEQ ID NO:59 or non-murine equivalents thereof, such as rat,human, chimeric or humanised variants thereof, for example they arederived from the humanised antibody having the heavy and light chains asdescribed in SEQ ID NO:22 and SEQ ID NO:24.

In one aspect of the invention there is provided an antigen bindingprotein, for example an antibody which binds human IL-13 and whichcomprises variants of the CDRH3 SIYDDYHYDDYYAMDY (SEQ ID NO: 3), whereinCDRH3 is substituted by the alternative amino acids set out below at oneor more of the following positions (using Kabat numbering):

-   -   a) S95 in position 1 is substituted for tryptophan (W)    -   b) 196 in position 2 is substituted for valine (V)    -   c) Y97 in position 3 is substituted for phenylalanine (F)    -   d) D98 in position 4 is substituted for glutamine (E)    -   e) H100A in position 7 is substituted for alanine (A), glutamic        acid (E), glutamine (Q), Arginine (R), Serine (S), threonine (T)        or valine (V), and    -   f) Y100B in position 8 is substituted for alanine (A),        isoleucine, (1), tryptophan (W) or valine (V).

In another aspect of the invention there is provided an antigen bindingprotein, for example an antibody which binds human IL-13 and whichcomprises the CDRH3 set out in SEQ ID NO: 3, wherein CDRH3 comprises oneor more of the following substitutions: S95W, 196V, Y97F, D98E, H100A_A,H100A_E, H100A_Q, H100A_R, H100A_S, H100A_T, H100A_V, Y100B_A, Y100B_I,Y100B_W, and Y100B_V.

In one aspect the antigen binding protein of the present invention, forexample the antibody of the present invention, comprises a CDRH3sequence selected from those set out in SEQ ID NO: 4, SEQ ID NO: 5, SEQID NO: 6, SEQ ID NO: 7, SEQ ID NO:8 and SEQ ID NO: 9, SEQ ID NO: 10, SEQID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14 and SEQ ID NO: 15,SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO: 18. Such antigen bindingproteins may further comprise the following CDR sequences:

CDRH1: selected from SEQ ID NO:1, 60 and 61,

CDRH2: SEQ ID NO: 2; CDRL1: SEQ ID NO:19; CDRL2: SEQ ID NO:20; andCDRL3: SEQ ID NO:21.

In one embodiment the antigen binding protein comprises the followingCDRs:

CDRH1: SEQ ID NO:1, CDRH2: SEQ ID NO:2; CDRH3: SEQ ID NO:18 CDRL1: SEQID NO:19; CDRL2: SEQ ID NO:20; and CDRL3: SEQ ID NO:21.

In another embodiment the antigen binding protein comprises thefollowing CDRs:

CDRH1: SEQ ID NO:1, CDRH2: SEQ ID NO:2; CDRH3: SEQ ID NO:17 CDRL1: SEQID NO:19; CDRL2: SEQ ID NO:20; and CDRL3: SEQ ID NO:21.

In another embodiment the antigen binding protein comprises thefollowing CDRs:

CDRH1: SEQ ID NO:1, CDRH2: SEQ ID NO:2; CDRH3: SEQ ID NO:16 CDRL1: SEQID NO:19; CDRL2: SEQ ID NO:20; and CDRL3: SEQ ID NO:21.

In another embodiment the antigen binding protein comprises thefollowing CDRs:

CDRH1: SEQ ID NO:1, CDRH2: SEQ ID NO:2; CDRH3: SEQ ID NO:15 CDRL1: SEQID NO:19; CDRL2: SEQ ID NO:20; and CDRL3: SEQ ID NO:21.

Throughout this specification, amino acid residues in antibody sequencesare numbered according to the Kabat scheme. Similarly, the terms “CDR”,“CDRL1”, “CDRL2”, “CDRL3”, “CDRH1”, “CDRH2”, “CDRH3”, unless otherwiseindicated (e.g. CDRH3 as set out in SEQ ID NO:60 and 61), follow theKabat numbering system as set forth in Kabat et al; “Sequences ofproteins of Immunological Interest” NIH, 1987.

In another aspect of the invention there is provided an antigen bindingprotein, such as a humanised antibody or antigen binding fragmentthereof, comprising a heavy chain having the sequence selected from SEQID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34,SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO:44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ IDNO: 54, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90 and SEQ ID NO: 91;and the light chain of SEQ ID NO:24.

The invention provides an antigen binding protein, such as a humanisedantibody or antigen binding fragment thereof, comprising a heavy chainselected from SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO:32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ IDNO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQID NO: 52, SEQ ID NO: 54, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90.

The invention also provides an antigen binding protein, such as ahumanised antibody or antigen binding fragment thereof, comprising alight chain selected from SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112and SEQ ID NO: 114.

The invention further provides an antigen binding protein, such as ahumanised antibody or antigen binding fragment thereof, comprising aheavy chain having the sequence selected from SEQ ID NO: 26, SEQ ID NO:28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ IDNO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 88,SEQ ID NO: 89, SEQ ID NO: 90 and SEQ ID NO: 91; and the light chain ofSEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112 and SEQ ID NO: 114.

In one embodiment the antigen binding protein of the present inventioncomprises an antibody comprising a heavy chain selected from SEQ ID NO:26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ IDNO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54,SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90 and SEQ ID NO: 91 and alight chain selected from SEQ ID NO:24, SEQ ID NO: 108, SEQ ID NO: 110,SEQ ID NO: 112 and SEQ ID NO: 114.

In one embodiment the antigen binding protein of the present inventioncomprises a heavy chain selected from SEQ ID NO: 48, SEQ ID NO: 50, SEQID NO: 52 and SEQ ID NO: 54; and a light chain selected from SEQ ID NO:24, SEQ ID NO: 108, SEQ ID NO:110, SEQ ID NO:112 and SEQ ID NO:114, forexample the antigen binding protein comprises the heavy chain of SEQ IDNO: 48 and the light chain of SEQ ID NO:24, or the heavy chain of SEQ IDNO: 50 and the light chain of SEQ ID NO:24, or the heavy chain of SEQ IDNO: 52 and the light chain of SEQ ID NO:24, or the heavy chain of SEQ IDNO: 54 and the light chain of SEQ ID NO:24, or the heavy chain of SEQ IDNO: 88 and the light chain of SEQ ID NO:24 or the heavy chain of SEQ IDNO: 89 and the light chain of SEQ ID NO:24, or the heavy chain of SEQ IDNO: 90 and the light chain of SEQ ID NO:24, or the heavy chain of SEQ IDNO: 91 and the light chain of SEQ ID NO:24, or the heavy chain of SEQ IDNO: 48 and the light chain of SEQ ID NO:108, or the heavy chain of SEQID NO: 50 and the light chain of SEQ ID NO:108, or the heavy chain ofSEQ ID NO: 52 and the light chain of SEQ ID NO:108, or the heavy chainof SEQ ID NO: 54 and the light chain of SEQ ID NO:108, or the heavychain of SEQ ID NO: 88 and the light chain of SEQ ID NO:108 or the heavychain of SEQ ID NO: 89 and the light chain of SEQ ID NO:108, or theheavy chain of SEQ ID NO: 90 and the light chain of SEQ ID NO:108, orthe heavy chain of SEQ ID NO: 91 and the light chain of SEQ ID NO:108,or the heavy chain of SEQ ID NO: 48 and the light chain of SEQ IDNO:110, or the heavy chain of SEQ ID NO: 50 and the light chain of SEQID NO:110, or the heavy chain of SEQ ID NO: 52 and the light chain ofSEQ ID NO:110, or the heavy chain of SEQ ID NO: 54 and the light chainof SEQ ID NO:110, or the heavy chain of SEQ ID NO: 88 and the lightchain of SEQ ID NO:110 or the heavy chain of SEQ ID NO: 89 and the lightchain of SEQ ID NO:110, or the heavy chain of SEQ ID NO: 90 and thelight chain of SEQ ID NO:110, or the heavy chain of SEQ ID NO: 91 andthe light chain of SEQ ID NO:110.

In one such embodiment the antigen binding protein of the presentinvention comprises a heavy chain selected from SEQ ID NO: 48, SEQ IDNO: 50, SEQ ID NO: 52 and SEQ ID NO: 54; and a light chain selected fromSEQ ID NO:108 and SEQ ID NO:110, for example the antigen binding proteincomprises the heavy chain of SEQ ID NO: 48 and the light chain of SEQ IDNO:108, or the antigen binding protein comprises the heavy chain of SEQID NO: 48 and the light chain of SEQ ID NO:110, or the antigen bindingprotein comprises the heavy chain of SEQ ID NO: 50 and the light chainof SEQ ID NO:108, or the antigen binding protein comprises the heavychain of SEQ ID NO: 50 and the light chain of SEQ ID NO:110, or theantigen binding protein comprises the heavy chain of SEQ ID NO: 52 andthe light chain of SEQ ID NO:108, or the antigen binding proteincomprises the heavy chain of SEQ ID NO: 52 and the light chain of SEQ IDNO:110, or the antigen binding protein comprises the heavy chain of SEQID NO: 54 and the light chain of SEQ ID NO:108, or the antigen bindingprotein comprises the heavy chain of SEQ ID NO: 54 and the light chainof SEQ ID NO:110.

The IL-13 antibodies of the present invention may be combined with anIL-4 and/or an IL-5 antagonist, for example an IL-4 antibody or epitopebinding domain, and/or an IL-5 antibody or epitope binding domain. Thesemay be administered as a mixture of separate molecules which areadministered at the same time i.e. co-administered, or are administeredwithin 24 hours of each other, for example within 20 hours, or within 15hours or within 12 hours, or within 10 hours, or within 8 hours, orwithin 6 hours, or within 4 hours, or within 2 hours, or within 1 hour,or within 30 minutes of each other.

In a further embodiment the antagonists are present as one moleculecapable of binding to two or more antigens, for example the inventionprovides an antigen binding protein comprising the IL-13 antibody of thepresent invention which is capable of binding to IL-13 and which is alsocapable of binding to IL-4 or which is also capable of binding to IL-5,or which is also capable of binding to IL-4 and IL-5.

In one embodiment the antigen binding protein of the present inventionmay be a multi-specific antibody which comprises at least CDRH3, andoptionally one or more of CDRH1, CDRH2, CDRL1, CDRL2, and CDRL3 of thepresent invention, which is capable of binding to IL-13 and which isalso capable of binding to one or more of IL-4 or IL-5. In one suchembodiment, a multi-specific antibody is provided which comprises aCDRH3, or an antigen binding protein as defined herein, and whichcomprises a further antigen binding site which is capable of binding toIL-4, or IL-5.

One example of an antigen binding protein of the present invention is anantibody specific for IL-13 comprising CDRH1, CDRH2, CDRH3, CDRL1,CDRL2, and CDRL3 as defined herein, linked to one or moreepitope-binding domains which have specificity for IL-4 or IL-5, forexample a bispecific antigen binding protein which is capable of bindingto IL-13 and IL-4, or IL-13 and IL-5, or a trispecific antigen bindingprotein which is capable of binding to IL-13, IL-4 and IL-5.

It will be understood that any of the antigen-binding proteins describedherein may be capable of binding two or more antigens simultaneously,for example, as determined by stochiometry analysis by using a suitableassay such as that described in Example 8.

The present invention provides an antigen-binding protein comprising theIL-13 antibody of the present invention which is linked to one or moreepitope-binding domains, for example an antigen-binding proteincomprising the IL-13 antibody of the present invention linked to anepitope-binding domain which is capable of binding to IL-4, or anantigen-binding protein comprising the IL-13 antibody of the presentinvention linked to an epitope-binding domain which is capable ofbinding to IL-5, or an antigen-binding protein comprising the IL-13antibody of the present invention linked to a first epitope-bindingdomain which is capable of binding to IL-4 and a second epitope-bindingdomain which is capable of binding to IL-5.

The epitope-binding domain may be attached to the c-terminus or then-terminus of the heavy chain of the IL-13 antibody or the c-terminus orn-terminus of the light chain of the IL-13 antibody.

Antibodies of the present invention may be linked to epitope bindingdomains by the use of linkers. Examples of suitable linkers includeamino acid sequences which may be from 1 amino acid to 150 amino acidsin length, or from 1 amino acid to 140 amino acids, for example, from 1amino acid to 130 amino acids, or from 1 to 120 amino acids, or from 1to 80 amino acids, or from 1 to 50 amino acids, or from 1 to 20 aminoacids, or from 1 to 10 amino acids, or from 5 to 18 amino acids. Suchsequences may have their own tertiary structure, for example, a linkerof the present invention may comprise a single variable domain. The sizeof a linker in one embodiment is equivalent to a single variable domain.Suitable linkers may be of a size from 1 to 20 angstroms, for exampleless than 15 angstroms, or less than 10 angstroms, or less than 5angstroms.

In one embodiment of the present invention at least one of the epitopebinding domains is directly attached to the IL-13 antibody with a linkercomprising from 1 to 150 amino acids, for example 1 to 20 amino acids,for example 1 to 10 amino acids.

Such linkers may be selected from any one of those set out in SEQ IDNO:82-87, 92 to 93. or multiples of such linkers.

Linkers of use in the antigen-binding proteins of the present inventionmay comprise alone or in addition to other linkers, one or more sets ofGS residues, for example ‘GSTVAAPS’ (SEQ ID NO: 92) or ‘TVAAPSGS’ (SEQID NO: 87) or ‘GSTVAAPSGS’ (SEQ ID NO: 93). In one embodiment the linkercomprises SEQ ID NO: 83.

In one embodiment the epitope binding domain is linked to the IL-13antibody by the linker ‘(PAS)_(n)(GS)_(m)’. In another embodiment theepitope binding domain is linked to the IL-13 antibody by the linker‘(GGGGS)_(n)(GS)_(m)’. In another embodiment the epitope binding domainis linked to the IL-13 antibody by the linker ‘(TVAAPS)_(n)(GS)_(m)’. Inanother embodiment the epitope binding domain is linked to the IL-13antibody by the linker ‘(GS)_(m)(TVAAPSGS)_(n)’. In another embodimentthe epitope binding domain is linked to the IL-13 antibody by the linker‘(PAVPPP)_(n)(GS)_(m)’. In another embodiment the epitope binding domainis linked to the IL-13 antibody by the linker ‘(TVSDVP)_(n)(GS)_(m)’. Inanother embodiment the epitope binding domain is linked to the IL-13antibody by the linker ‘(TGLDSP)_(n)(GS)_(m)’. In all such embodiments,n=1-10, and m=0-4.

Examples of such linkers include (PAS)_(n)(GS)_(m) wherein n=1 and m=1,(PAS)_(n)(GS)_(m) wherein n=2 and m=1, (PAS)_(n)(GS)_(m) wherein n=3 andm=1, (PAS)_(n)(GS)_(m) wherein n=4 and m=1, (PAS)_(n)(GS)_(m) whereinn=2 and m=0, (PAS)_(n)(GS)_(m) wherein n=3 and m=0, (PAS)_(n)(GS)_(m)wherein n=4 and m=0.

Examples of such linkers include (GGGGS)_(n)(GS)_(m) wherein n=1 andm=1, (GGGGS)_(n)(GS)_(m) wherein n=2 and m=1, (GGGGS)_(n)(GS)_(m)wherein n=3 and m=1, (GGGGS)_(n)(GS)_(m) wherein n=4 and m=1,(GGGGS)_(n)(GS)_(m) wherein n=2 and m=0, (GGGGS)_(n)(GS)_(m) wherein n=3and m=0, (GGGGS)_(n)(GS)_(m) wherein n=4 and m=0.

Examples of such linkers include (TVAAPS)_(n)(GS)_(m) wherein n=1 andm=1 (SEQ ID NO:87), (TVAAPS)_(n)(GS)_(m) wherein n=2 and m=1 (SEQ IDNO:145), (TVAAPS)_(n)(GS)_(m) wherein n=3 and m=1 (SEQ ID NO:146),(TVAAPS)_(n)(GS)_(m) wherein n=4 and m=1, (TVAAPS)_(n)(GS)_(m) whereinn=2 and m=0, (TVAAPS)_(n)(GS)_(m) wherein n=3 and m=0,(TVAAPS)_(n)(GS)_(m) wherein n=4 and m=0.

Examples of such linkers include (GS)_(m)(TVAAPSGS)_(n) wherein n=1 andm=1 (SEQ ID NO:139), (GS)_(m)(TVAAPSGS)_(n) wherein n=2 and m=1 (SEQ IDNO:140), (GS)_(m)(TVAAPSGS)_(n) wherein n=3 and m=1 (SEQ ID NO:141), or(GS)_(m)(TVAAPSGS)_(n) wherein n=4 and m=1 (SEQ ID NO:142),(GS)_(m)(TVAAPSGS)_(n) wherein n=5 and m=1 (SEQ ID NO:143),(GS)_(m)(TVAAPSGS)_(n) wherein n=6 and m=1 (SEQ ID NO:144),(GS)_(m)(TVAAPSGS)_(n) wherein n=1 and m=0 (SEQ ID NO:87),(GS)_(m)(TVAAPSGS)_(n) wherein n=2 and m=10, (GS)_(m)(TVAAPSGS)_(n)wherein n=3 and m=0, or (GS)_(m)(TVAAPSGS)_(n) wherein n=0.

Examples of such linkers include (PAVPPP)_(n)(GS)_(m) wherein n=1 andm=1, (PAVPPP)_(n)(GS)_(m) wherein n=2 and m=1 (SEQ ID NO: 65),(PAVPPP)_(n)(GS)_(m) wherein n=3 and m=1, (PAVPPP)_(n)(GS)_(m) whereinn=4 and m=1, (PAVPPP)_(n)(GS)_(m) wherein n=2 and m=0,(PAVPPP)_(n)(GS)_(m) wherein n=3 and m=0, (PAVPPP)_(n)(GS)_(m) whereinn=4 and m=0.

Examples of such linkers include (TVSDVP)_(n)(GS)_(m) wherein n=1 andm=1 (SEQ ID NO: 67), (TVSDVP)_(n)(GS)_(m) wherein n=2 and m=1,(TVSDVP)_(n)(GS)_(m) wherein n=3 and m=1, (TVSDVP)_(n)(GS)_(m) whereinn=4 and m=1, (TVSDVP)_(n)(GS)_(m) wherein n=2 and m=0,(TVSDVP)_(n)(GS)_(m) wherein n=3 and m=0, (TVSDVP)_(n)(GS)_(m) whereinn=4 and m=0.

Examples of such linkers include (TGLDSP)_(n)(GS)_(m) wherein n=1 andm=1, (TGLDSP)_(n)(GS)_(m) wherein n=2 and m=1, (TGLDSP)_(n)(GS)_(m)wherein n=3 and m=1, (TGLDSP)_(n)(GS)_(m) wherein n=4 and m=1,(TGLDSP)_(n)(GS)_(m) wherein n=2 and m=0, (TGLDSP)_(n)(GS)_(m) whereinn=3 and m=0, (TGLDSP)_(n)(GS)_(m) wherein n=4 and m=0.

In another embodiment there is no linker between the epitope bindingdomain and the IL-13 antibody. In another embodiment the epitope bindingdomain is linked to the IL-13 antibody by the linker TVAAPS' (SEQ ID NO:83). In another embodiment the epitope binding domain, is linked to theIL-13 antibody by the linker TVAAPSGS' (SEQ ID NO: 87). In anotherembodiment the epitope binding domain is linked to the IL-13 antibody bythe linker ‘GS’. In another embodiment the epitope binding domain islinked to the IL-13 antibody by the linker ‘ASTKGPT’ (SEQ ID NO: 84).

Epitope-binding domains of use in the present invention are domains thatspecifically bind an antigen or epitope independently of a different Vregion or domain, this may be a domain antibody or may be anon-Immunoglobulin domain, for example a domain which is a derivative ofa scaffold selected from the group consisting of CTLA-4 (Evibody);lipocalin; Protein A derived molecules such as Z-domain of Protein A(Affibody, SpA), A-domain (Avimer/Maxibody); Heat shock proteins such asGroEI and GroES; transferrin (trans-body); ankyrin repeat protein(DARPin); peptide aptamer; C-type lectin domain (Tetranectin); humanγ-crystallin and human ubiquitin (affilins); PDZ domains; scorpiontoxinkunitz type domains of human protease inhibitors; and fibronectin(adnectin); which has been subjected to protein engineering in order toobtain binding to a ligand other than the natural ligand. In oneembodiment this may be an domain antibody or other suitable domains suchas a domain selected from the group consisting of CTLA-4, lipocallin,SpA, an Affibody, an avimer, GroEI, transferrin, GroES and fibronectin.In one embodiment this may be selected from an immunoglobulin singlevariable domain, an Affibody, an ankyrin repeat protein (DARPin) and anadnectin. In another embodiment this may be selected from an Affibody,an ankyrin repeat protein (DARPin) and an adnectin. In anotherembodiment this may be a domain antibody, for example a domain antibodyselected from a human, camelid (nanobody), or shark (NARV) domainantibody.

Examples of such antigen-binding proteins include the IL-13 antibodiesof the present invention which have an epitope binding domain which isan IL-4 antagonist attached to the c-terminus or the n-terminus of theheavy chain or the c-terminus. Examples include an antigen bindingprotein comprising the heavy chain sequence set out in SEQ ID NO: 26,SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO:36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ IDNO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQID NO: 98, SEQ ID NO: 100, SEQ ID NO:102, SEQ ID NO:104, SEQ ID NO:106or SEQ ID NO: 117-138, and the light chain sequence set out in SEQ IDNO: 24, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112 or SEQ ID NO:114, wherein one or both of the Heavy and Light chain further compriseone or more epitope-binding domains which is capable of antagonisingIL-4, for example single variable domains which are capable of bindingto IL-4. Such epitope-binding domains can be selected from those set outin SEQ ID NO: 78-81 and 94.

In one embodiment the antigen binding constructs of the presentinvention comprise the heavy chain sequence of SEQ ID NO: 62 and thelight chain sequence of SEQ ID NO: 24, or the heavy chain sequence ofSEQ ID NO: 64 and the light chain sequence of SEQ ID NO: 24, or theheavy chain sequence of SEQ ID NO: 66 and the light chain sequence ofSEQ ID NO: 24, or the heavy chain sequence of SEQ ID NO: 68 and thelight chain sequence of SEQ ID NO: 24, or the heavy chain sequence ofSEQ ID NO: 70 and the light chain sequence of SEQ ID NO: 24, or theheavy chain sequence of SEQ ID NO: 72 and the light chain sequence ofSEQ ID NO: 24, or the heavy chain sequence of SEQ ID NO:74 and the lightchain sequence of SEQ ID NO: 24, or the heavy chain sequence of SEQ IDNO: 76 and the light chain sequence of SEQ ID NO: 24.

In one embodiment the antigen binding constructs of the presentinvention comprise the heavy chain sequence of SEQ ID NO: 94 and thelight chain sequence of SEQ ID NO: 24, or the heavy chain sequence ofSEQ ID NO: 96 and the light chain sequence of SEQ ID NO: 24, or theheavy chain sequence of SEQ ID NO: 98 and the light chain sequence ofSEQ ID NO: 24, or the heavy chain sequence of SEQ ID NO: 100 and thelight chain sequence of SEQ ID NO: 24, or the heavy chain sequence ofSEQ ID NO: 102 and the light chain sequence of SEQ ID NO: 24, or theheavy chain sequence of SEQ ID NO: 104 and the light chain sequence ofSEQ ID NO: 24, or the heavy chain sequence of SEQ ID NO:106 and thelight chain sequence of SEQ ID NO: 24.

In one embodiment the antigen binding constructs of the presentinvention comprise the heavy chain sequence of SEQ ID NO: 62 and thelight chain sequence of SEQ ID NO: 108, or the heavy chain sequence ofSEQ ID NO: 64 and the light chain sequence of SEQ ID NO: 108, or theheavy chain sequence of SEQ ID NO: 66 and the light chain sequence ofSEQ ID NO: 108, or the heavy chain sequence of SEQ ID NO: 68 and thelight chain sequence of SEQ ID NO: 108, or the heavy chain sequence ofSEQ ID NO: 70 and the light chain sequence of SEQ ID NO: 108, or theheavy chain sequence of SEQ ID NO: 72 and the light chain sequence ofSEQ ID NO: 108, or the heavy chain sequence of SEQ ID NO:74 and thelight chain sequence of SEQ ID NO: 108, or the heavy chain sequence ofSEQ ID NO: 76 and the light chain sequence of SEQ ID NO: 108, or theheavy chain sequence of SEQ ID NO: 62 and the light chain sequence ofSEQ ID NO: 110, or the heavy chain sequence of SEQ ID NO: 64 and thelight chain sequence of SEQ ID NO: 110, or the heavy chain sequence ofSEQ ID NO: 66 and the light chain sequence of SEQ ID NO: 110, or theheavy chain sequence of SEQ ID NO: 68 and the light chain sequence ofSEQ ID NO: 110, or the heavy chain sequence of SEQ ID NO: 70 and thelight chain sequence of SEQ ID NO: 110, or the heavy chain sequence ofSEQ ID NO: 72 and the light chain sequence of SEQ ID NO: 110, or theheavy chain sequence of SEQ ID NO:74 and the light chain sequence of SEQID NO: 110, or the heavy chain sequence of SEQ ID NO: 76 and the lightchain sequence of SEQ ID NO: 110.

In one embodiment the antigen binding constructs of the presentinvention comprise the heavy chain sequence of SEQ ID NO: 96 and thelight chain sequence of SEQ ID NO: 108, or the heavy chain sequence ofSEQ ID NO: 98 and the light chain sequence of SEQ ID NO: 108, or theheavy chain sequence of SEQ ID NO: 100 and the light chain sequence ofSEQ ID NO: 108, or the heavy chain sequence of SEQ ID NO: 102 and thelight chain sequence of SEQ ID NO: 108, or the heavy chain sequence ofSEQ ID NO: 104 and the light chain sequence of SEQ ID NO: 108, or theheavy chain sequence of SEQ ID NO: 106 and the light chain sequence ofSEQ ID NO: 108, or the heavy chain sequence of SEQ ID NO: 96 and thelight chain sequence of SEQ ID NO: 110, or the heavy chain sequence ofSEQ ID NO: 98 and the light chain sequence of SEQ ID NO: 110, or theheavy chain sequence of SEQ ID NO: 100 and the light chain sequence ofSEQ ID NO: 110, or the heavy chain sequence of SEQ ID NO: 102 and thelight chain sequence of SEQ ID NO: 110, or the heavy chain sequence ofSEQ ID NO: 104 and the light chain sequence of SEQ ID NO: 110, or theheavy chain sequence of SEQ ID NO: 106 and the light chain sequence ofSEQ ID NO: 110.

In one embodiment the IL-13 antibody heavy chain is selected from thoseset out in SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52 and SEQ ID NO:54. In another embodiment the heavy chain is selected from those set outin SEQ ID NO:88-91, SEQ ID NO:96-106, and SEQ ID NO:117-138. In one suchembodiment the heavy chain is selected from SEQ ID NO:96, SEQ ID NO: 98,SEQ ID NO:100, SEQ ID NO:102, SEQ ID NO:104 and SEQ ID NO:106.

In one embodiment the antigen-binding protein will comprise ananti-IL-13 antibody linked to an epitope binding domain which is a IL-5antagonist, wherein the anti-IL-13 antibody comprises the CDRH3 selectedfrom those set out in SEQ ID NO: 3-18, for example SEQ ID NO: 15-18 andthe light chain sequence of SEQ ID NO: 24.

Examples include an antigen binding protein comprising the heavy chainsequence set out in SEQ ID NO: 26, 28, 30, 32, 34, 36, 38, 40, 42, 44,46, 48, 50, 52 or 54 and the light chain sequence set out in SEQ ID NO:24 wherein one or both of the Heavy and Light chain further comprise oneor more epitope-binding domains which is capable of antagonising IL-5,for example immunoglobulin single variable domains which are capable ofbinding to IL-5.

In a further embodiment, the antigen binding protein will comprise theheavy chain sequence set out in SEQ ID NO: 26, 28, 30, 32, 34, 36, 38,40, 42, 44, 46, 48, 50, 52 or 54 and the light chain sequence set out inSEQ ID NO: 24 wherein one or both of the Heavy and Light chain furthercomprise one or more epitope-binding domains which are capable ofantagonising IL-4 for example immunoglobulin single variable domainswhich are capable of binding to IL-4, and one or more epitope-bindingdomains which are capable of antagonising IL-5, for exampleimmunoglobulin single variable domains which are capable of binding toIL-5.

In one embodiment, the antigen-binding protein of the present inventioncomprises at least one epitope binding domain, which is capable ofbinding human serum albumin.

In one embodiment, there are at least 3 antigen-binding sites, forexample there are 4, or 5 or 6 or 8 or 10 antigen-binding sites and theantigen-binding protein is capable of binding at least 3 or 4 or 5 or 6or 8 or 10 antigens, for example it is capable of binding 3 or 4 or 5 or6 or 8 or 10 antigens simultaneously.

In one embodiment, a first epitope binding domain is linked to theprotein scaffold and a second epitope binding domain is linked to thefirst epitope binding domain, for example where the protein scaffold isan IgG scaffold, a first epitope binding domain may be linked to thec-terminus of the heavy chain of the IgG scaffold, and that epitopebinding domain can be linked at its c-terminus to a second epitopebinding domain, or for example a first epitope binding domain may belinked to the c-terminus of the light chain of the IgG scaffold, andthat first epitope binding domain may be further linked at itsc-terminus to a second epitope binding domain, or for example a firstepitope binding domain may be linked to the n-terminus of the lightchain of the IgG scaffold, and that first epitope binding domain may befurther linked at its n-terminus to a second epitope binding domain, orfor example a first epitope binding domain may be linked to then-terminus of the heavy chain of the IgG scaffold, and that firstepitope binding domain may be further linked at its n-terminus to asecond epitope binding domain.

When the epitope-binding domain is a domain antibody, some domainantibodies may be suited to particular positions within the scaffold.

Immunoglobulin single variable domains of use in the present inventioncan be linked at the C-terminal end of the heavy chain and/or the lightchain of the IL-13 mAb. In addition some immunoglobulin single variabledomains can be linked to the C-terminal ends of both the heavy chain andthe light chain of conventional antibodies.

In constructs where the N-terminus of immunoglobulin single variabledomains are fused to an antibody constant domain (either C_(H)3 or CL),a peptide linker may help the immunoglobulin single variable domain tobind to antigen. Indeed, the N-terminal end of a dAb is located closelyto the CDRs involved in antigen-binding activity. Thus a short peptidelinker acts as a spacer between the epitope-binding domain, and theconstant domain of the antibody, which may allow the dAb CDRs to moreeasily reach the antigen, which may therefore bind with high affinity.

The surroundings in which immunoglobulin single variable domains arelinked to the IgG will differ depending on which antibody chain they arefused to: When fused at the C-terminal end of the antibody light chain,each immunoglobulin single variable domain is expected to be located inthe vicinity of the antibody hinge and the Fc portion. It is likely thatsuch immunoglobulin single variable domains will be located far apartfrom each other. In conventional antibodies, the angle between Fabfragments and the angle between each Fab fragment and the Fc portion canvary quite significantly. It is likely that—with mAbdAbs—the anglebetween the Fab fragments will not be widely different, whilst someangular restrictions may be observed with the angle between each Fabfragment and the Fc portion.

When fused at the C-terminal end of the antibody heavy chain, eachimmunoglobulin single variable domain is expected to be located in thevicinity of the C_(H)3 domains of the Fc portion. This is not expectedto impact on the Fc binding properties to Fc receptors (e.g. FcγRI, II,III an FcRn) as these receptors engage with the C_(H)2 domains (for theFcγRI, II and III class of receptors) or with the hinge between theC_(H)2 and C_(H)3 domains (e.g. FcRn receptor). Another feature of suchantigen-binding proteins is that both immunoglobulin single variabledomains are expected to be spatially close to each other and providedthat flexibility is provided by provision of appropriate linkers, theseimmunoglobulin single variable domains may even form homodimericspecies, hence propagating the ‘zipped’ quaternary structure of the Fcportion, which may enhance stability of the construct.

Such structural considerations can aid in the choice of the mostsuitable position to link an epitope-binding domain, for example animmunoglobulin single variable domain, on to an antibody.

The size of the antigen, its localization (in blood or on cell surface),its quaternary structure (monomeric or multimeric) can vary.Conventional antibodies are naturally designed to function as adaptorconstructs due to the presence of the hinge region, wherein theorientation of the two antigen-binding sites at the tip of the Fabfragments can vary widely and hence adapt to the molecular feature ofthe antigen and its surroundings. In contrast immunoglobulin singlevariable domains linked to an antibody with no hinge region, may haveless structural flexibility either directly or indirectly.

Understanding the solution state and mode of binding at theimmunoglobulin single variable domain is also helpful. Evidence hasaccumulated that in vitro human dAbs can predominantly exist inmonomeric, homo-dimeric or multimeric forms in solution (Reiter et al.(1999) J Mol Biol 290 p685-698; Ewert et al (2003) J Mol Biol 325,p531-553, Jespers et al (2004) J Mol Biol 337 p893-903; Jespers et al(2004) Nat Biotechnol 22 p1161-1165; Martin et al (1997) Protein Eng. 10p607-614; Sepulvada et al (2003) J Mol Biol 333 p355-365). This isfairly reminiscent to multimerisation events observed in vivo with Igdomains such as Bence-Jones proteins (which are dimers of immunoglobulinlight chains (Epp et al (1975) Biochemistry 14 p4943-4952; Huan et al(1994) Biochemistry 33 p14848-14857; Huang et al (1997) Mol immunol 34p1291-1301) and amyloid fibers (James et al. (2007) J Mol. Biol.367:603-8).

For example, it may be desirable to link dAbs that tend to dimerise insolution to the C-terminal end of the Fc portion in preference to theC-terminal end of the light chain as linking to the C-terminal end ofthe Fc will allow those dAbs to dimerise in the context of theantigen-binding protein of the invention.

The antigen-binding proteins of the present invention may compriseantigen-binding sites specific for a single antigen, or may haveantigen-binding sites specific for two or more antigens, or for two ormore epitopes on a single antigen, or there may be antigen-binding siteseach of which is specific for a different epitope on the same ordifferent antigens.

The antigen binding proteins of the invention may comprise heavy chainvariable regions and light chain variable regions of the invention whichmay be formatted into the structure of a natural antibody or functionalfragment or equivalent thereof. An antigen binding protein of theinvention may therefore comprise the VH regions of the inventionformatted into a full length antibody, a (Fab′)₂ fragment, a Fabfragment, or equivalent thereof (such as scFV, bi- tri- or tetra-bodies,Tandabs etc.), when paired with an appropriate light chain. The antibodymay be an IgG1, IgG2, IgG3, or IgG4; or IgM; IgA, IgE or IgD or amodified variant thereof. The constant domain of the antibody heavychain may be selected accordingly. The light chain constant domain maybe a kappa or lambda constant domain. Furthermore, the antigen bindingprotein may comprise modifications of all classes e.g. IgG dimers, Fcmutants that no longer bind Fc receptors or mediate Clq binding. Theantigen binding protein may also be a chimeric antibody of the typedescribed in WO86/01533 which comprises an antigen binding region and anon-immunoglobulin region.

The constant region is selected according to any functionality required.An IgG1 may demonstrate lytic ability through binding to complementand/or will mediate ADCC (antibody dependent cell cytotoxicity). An IgG4can be used if a non-cytotoxic blocking antibody is required. However,IgG4 antibodies can demonstrate instability in production and thereforean alternative is to modify the generally more stable IgG1. Suggestedmodifications are described in EP0307434, for example mutations atpositions 235 and 237. The invention therefore provides a lytic or anon-lytic form of an antigen binding protein, for example an antibodyaccording to the invention.

In certain forms the antibody of the invention is a full length (e.g.H2L2 tetramer) lytic or non-lytic IgG1 antibody having any of the heavychain variable regions described herein. In a further aspect, theinvention provides polynucleotides encoding the light and heavy chainvariable regions as described herein.

In one embodiment of the invention the antigen-binding site binds toantigen with a Kd of at least about 1 mM, for example a Kd of at leastabout 10 nM, at least about 1 nM, at least about 500 pM, at least about200 pM, at least about 100 pM, or at least about 50 pM to each antigenas measured by Biacore™.

In one embodiment of the invention the antigen-binding site binds toantigen with a Kd of at least about 1 mM, for example a Kd of at leastabout 10 nM, at least about 1 nM, at least about 500 pM, at least about200 pM, at least about 100 pM, or at least about 50 pM to each antigenas measured by Biacore™.

In one embodiment the invention provides antigen binding proteins whichhave at least a 2 fold higher affinity, or at least 4 fold higheraffinity, or at least 6 fold higher affinity, or at least 8 fold higheraffinity, or at least 10 fold higher affinity for human IL-13 asmeasured by Biacore than the anti-IL=13 antibody comprising the heavychain sequence set out in SEQ ID NO:22 and the light chain sequence setout in SEQ ID NO:24.

The term “neutralises” and grammatical variations thereof as usedthroughout the present specification in relation to antigen bindingproteins of the invention means that a biological activity of IL-13 isreduced, either totally or partially, in the presence of the antigenbinding proteins of the present invention in comparison to the activityof IL-13 in the absence of such antigen binding proteins. Neutralisationmay be due to but not limited to one or more of blocking ligand binding,preventing the ligand activating the receptor, down regulating the IL-13receptor or affecting effector functionality. Levels of neutralisationcan be measured in several ways, for example by use of the assays as setout in the examples below, for example in a TF1 assay which may becarried out for example as described in Example 4. The neutralisation ofIL-13, IL-4 or both of these cytokines in this assay is measured byassessing the inhibition of TF1 cell proliferation in the presence ofneutralising antigen binding protein.

Other methods of assessing neutralisation, for example, by assessing thedecreased binding between the IL-13 and its receptor in the presence ofneutralising antigen binding protein are known in the art, and include,for example, Biacore assays.

In an alternative aspect of the present invention there is providedantigen binding proteins which have at least substantially equivalentneutralising activity to the antibodies exemplified herein, for exampleantigen binding proteins which retain the neutralising activity ofA1Y100BAlaL1, A1Y100BIleL1, A1 Y100BTrpL1 or A1Y100BValL1 in a TF1 cellproliferation assay which can be carried out as set out in Example 4.

The antigen binding proteins, for example antibodies of the presentinvention may be produced by transfection of a host cell with anexpression vector comprising the coding sequence for the antigen bindingprotein of the invention. An expression vector or recombinant plasmid isproduced by placing these coding sequences for the antigen bindingprotein in operative association with conventional regulatory controlsequences capable of controlling the replication and expression in,and/or secretion from, a host cell. Regulatory sequences includepromoter sequences, e.g., CMV promoter, and signal sequences which canbe derived from other known antibodies. Similarly, a second expressionvector can be produced having a DNA sequence which encodes acomplementary antigen binding protein light or heavy chain. In certainembodiments this second expression vector is identical to the firstexcept insofar as the coding sequences and selectable markers areconcerned, so to ensure as far as possible that each polypeptide chainis functionally expressed. Alternatively, the heavy and light chaincoding sequences for the antigen binding protein may reside on a singlevector. A selected host cell is co-transfected by conventionaltechniques with both the first and second vectors (or simply transfectedby a single vector) to create the transfected host cell of the inventioncomprising both the recombinant or synthetic light and heavy chains.

The transfected cell is then cultured by conventional techniques toproduce the engineered antigen binding protein of the invention. Theantigen binding protein which includes the association of both therecombinant heavy chain and/or light chain is screened from culture byappropriate assay, such as ELISA or RIA. Similar conventional techniquesmay be employed to construct other antigen binding proteins.

Suitable vectors for the cloning and subcloning steps employed in themethods and construction of the compositions of this invention may beselected by one of skill in the art. For example, the conventional pUCseries of cloning vectors may be used. One vector, pUC19, iscommercially available from supply houses, such as Amersham(Buckinghamshire, United Kingdom) or Pharmacia (Uppsala, Sweden).Additionally, any vector which is capable of replicating readily, has anabundance of cloning sites and selectable genes (e.g., antibioticresistance), and is easily manipulated may be used for cloning. Thus,the selection of the cloning vector is not a limiting factor in thisinvention.

The expression vectors may also be characterized by genes suitable foramplifying expression of the heterologous DNA sequences, e.g., themammalian dihydrofolate reductase gene (DHFR). Other vector sequencesinclude a poly A signal sequence, such as from bovine growth hormone(BGH) and the betaglobin promoter sequence (betaglopro). The expressionvectors useful herein may be synthesized by techniques well known tothose skilled in this art.

The components of such vectors, e.g. replicons, selection genes,enhancers, promoters, signal sequences and the like, may be obtainedfrom commercial or natural sources or synthesized by known proceduresfor use in directing the expression and/or secretion of the product ofthe recombinant DNA in a selected host. Other appropriate expressionvectors of which numerous types are known in the art for mammalian,bacterial, insect, yeast, and fungal expression may also be selected forthis purpose.

The present invention also encompasses a cell line transfected with arecombinant plasmid containing the coding sequences of the antigenbinding proteins of the present invention. Host cells useful for thecloning and other manipulations of these cloning vectors are alsoconventional. However, cells from various strains of E. coli may be usedfor replication of the cloning vectors and other steps in theconstruction of antigen binding proteins of this invention.

Suitable host cells or cell lines for the expression of the antigenbinding proteins of the invention include mammalian cells such as NSO,Sp2/0, CHO (e.g. DG44), COS, HEK, a fibroblast cell (e.g., 3T3), andmyeloma cells, for example it may be expressed in a CHO or a myelomacell. Human cells may be used, thus enabling the molecule to be modifiedwith human glycosylation patterns. Alternatively, other eukaryotic celllines may be employed. The selection of suitable mammalian host cellsand methods for transformation, culture, amplification, screening andproduct production and purification are known in the art. See, e.g.,Sambrook et al., cited above.

Bacterial cells may prove useful as host cells suitable for theexpression of the recombinant Fabs or other embodiments of the presentinvention (see, e.g., Plückthun, A., Immunol. Rev., 130:151-188 (1992)).However, due to the tendency of proteins expressed in bacterial cells tobe in an unfolded or improperly folded form or in a non-glycosylatedform, any recombinant Fab produced in a bacterial cell would have to bescreened for retention of antigen binding ability. If the moleculeexpressed by the bacterial cell was produced in a properly folded form,that bacterial cell would be a desirable host, or in alternativeembodiments the molecule may express in the bacterial host and then besubsequently re-folded. For example, various strains of E. coli used forexpression are well-known as host cells in the field of biotechnology.Various strains of B. subtilis, Streptomyces, other bacilli and the likemay also be employed in this method.

Where desired, strains of yeast cells known to those skilled in the artare also available as host cells, as well as insect cells, e.g.Drosophila and Lepidoptera and viral expression systems. See, e.g.Miller et al., Genetic Engineering, 8:277-298, Plenum Press (1986) andreferences cited therein.

The general methods by which the vectors may be constructed, thetransfection methods required to produce the host cells of theinvention, and culture methods necessary to produce the antigen bindingprotein of the invention from such host cell may all be conventionaltechniques. Typically, the culture method of the present invention is aserum-free culture method, usually by culturing cells serum-free insuspension. Likewise, once produced, the antigen binding proteins of theinvention may be purified from the cell culture contents according tostandard procedures of the art, including ammonium sulfateprecipitation, affinity columns, column chromatography, gelelectrophoresis and the like. Such techniques are within the skill ofthe art and do not limit this invention. For example, preparation ofaltered antibodies are described in WO 99/58679 and WO 96/16990.

Yet another method of expression of the antigen binding proteins mayutilize expression in a transgenic animal, such as described in U.S.Pat. No. 4,873,316. This relates to an expression system using theanimal's casein promoter which when transgenically incorporated into amammal permits the female to produce the desired recombinant protein inits milk.

In a further aspect of the invention there is provided a method ofproducing an antibody of the invention which method comprises the stepof culturing a host cell transformed or transfected with a vectorencoding the light and/or heavy chain of the antibody of the inventionand recovering the antibody thereby produced.

In accordance with the present invention there is provided a method ofproducing an anti-IL-13 antibody of the present invention which binds toand neutralises the activity of human IL-13 which method comprises thesteps of;

-   -   (a) providing a first vector encoding a heavy chain of the        antibody;    -   (b) providing a second vector encoding a light chain of the        antibody;    -   (c) transforming a mammalian host cell (e.g. CHO) with said        first and second vectors;    -   (d) culturing the host cell of step (c) under conditions        conducive to the secretion of the antibody from said host cell        into said culture media;    -   (e) recovering the secreted antibody of step (d).

Once expressed by the desired method, the antibody is then examined forin vitro activity by use of an appropriate assay. Presently conventionalELISA assay formats are employed to assess qualitative and quantitativebinding of the antibody to IL-13. Additionally, other in vitro assaysmay also be used to verify neutralizing efficacy prior to subsequenthuman clinical studies performed to evaluate the persistence of theantibody in the body despite the usual clearance mechanisms.

The dose and duration of treatment relates to the relative duration ofthe molecules of the present invention in the human circulation, and canbe adjusted by one of skill in the art depending upon the conditionbeing treated and the general health of the patient. It is envisagedthat repeated dosing (e.g. once a week or once every two weeks) over anextended time period (e.g. four to six months) maybe required to achievemaximal therapeutic efficacy.

The mode of administration of the therapeutic agent of the invention maybe any suitable route which delivers the agent to the host. The antigenbinding proteins, and pharmaceutical compositions of the invention areparticularly useful for parenteral administration, i.e., subcutaneously(s.c.), intrathecally, intraperitoneally, intramuscularly (i.m.),intravenously (i.v.), or intranasally. Therapeutic agents of theinvention may be prepared as pharmaceutical compositions containing aneffective amount of the antigen binding protein of the invention as anactive ingredient in a pharmaceutically acceptable carrier. In oneembodiment the prophylactic agent of the invention is an aqueoussuspension or solution containing the antigen binding protein in a formready for injection. In one embodiment the suspension or solution isbuffered at physiological pH. In one embodiment the compositions forparenteral administration will comprise a solution of the antigenbinding protein of the invention or a cocktail thereof dissolved in apharmaceutically acceptable carrier. In one embodiment the carrier is anaqueous carrier. A variety of aqueous carriers may be employed, e.g.,0.9% saline, 0.3% glycine, and the like. These solutions may be madesterile and generally free of particulate matter. These solutions may besterilized by conventional, well known sterilization techniques (e.g.,filtration). The compositions may contain pharmaceutically acceptableauxiliary substances as required to approximate physiological conditionssuch as pH adjusting and buffering agents, etc. The concentration of theantigen binding protein of the invention in such pharmaceuticalformulation can vary widely, i.e., from less than about 0.5%, usually ator at least about 1% to as much as about 15 or 20% by weight and will beselected primarily based on fluid volumes, viscosities, etc., accordingto the particular mode of administration selected.

Thus, a pharmaceutical composition of the invention for intramuscularinjection could be prepared to contain about 1 mL sterile bufferedwater, and between about 1 ng to about 100 mg, e.g. about 50 ng to about30 mg or about 5 mg to about 25 mg, of an antigen binding protein, forexample an antibody of the invention. Similarly, a pharmaceuticalcomposition of the invention for intravenous infusion could be made upto contain about 250 ml of sterile Ringer's solution, and about 1 toabout 30 or 5 mg to about 25 mg of an antigen binding protein of theinvention per ml of Ringer's solution. Actual methods for preparingparenterally administrable compositions are well known or will beapparent to those skilled in the art and are described in more detailin, for example, Remington's Pharmaceutical Science, 15th ed., MackPublishing Company, Easton, Pa.

For the preparation of intravenously administrable antigen bindingprotein formulations of the invention see Lasmar U and Parkins D “Theformulation of Biopharmaceutical products”, Pharma. Sci.Tech.today, page129-137, Vol. 3 (3 Apr. 2000); Wang, W “Instability, stabilisation andformulation of liquid protein pharmaceuticals”, Int. J. Pharm 185 (1999)129-188; Stability of Protein Pharmaceuticals Part A and B ed Ahern T.J., Manning M. C., New York, N.Y.: Plenum Press (1992); Akers, M. J.“Excipient-Drug interactions in Parenteral Formulations”, J. Pharm Sci91 (2002) 2283-2300; Imamura, K et al “Effects of types of sugar onstabilization of Protein in the dried state”, J Pharm Sci 92 (2003)266-274; Izutsu, Kkojima, S. “Excipient crystallinity and itsprotein-structure-stabilizing effect during freeze-drying”, J. Pharm.Pharmacol, 54 (2002) 1033-1039; Johnson, R, “Mannitol-sucrosemixtures-versatile formulations for protein lyophilization”, J. Pharm.Sci, 91 (2002) 914-922; and Ha, E Wang W, Wang Y. j. “Peroxide formationin polysorbate 80 and protein stability”, J. Pharm Sci, 91, 2252-2264,(2002) the entire contents of which are incorporated herein by referenceand to which the reader is specifically referred.

In one embodiment the therapeutic agent of the invention, when in apharmaceutical preparation, is present in unit dose forms. Theappropriate therapeutically effective dose will be determined readily bythose of skill in the art. Suitable doses may be calculated for patientsaccording to their weight, for example suitable doses may be in therange of about 0.1 to about 20 mg/kg, for example about 1 to about 20mg/kg, for example about 10 to about 20 mg/kg or for example about 1 toabout 15 mg/kg, for example about 10 to about 15 mg/kg. To effectivelytreat conditions such as asthma or IPF in a human, suitable doses may bewithin the range of about 0.1 to about 1000 mg, for example about 0.1 toabout 500 mg, for example about 500 mg, for example about 0.1 to about100 mg, or about 0.1 to about 80 mg, or about 0.1 to about 60 mg, orabout 0.1 to about 40 mg, or for example about 1 to about 100 mg, orabout 1 to about 50 mg, of an antigen binding protein of this invention,which may be administered parenterally, for example subcutaneously,intravenously or intramuscularly. Such dose may, if necessary, berepeated at appropriate time intervals selected as appropriate by aphysician.

The antigen binding proteins described herein can be lyophilized forstorage and reconstituted in a suitable carrier prior to use. Thistechnique has been shown to be effective with conventionalimmunoglobulins and art-known lyophilization and reconstitutiontechniques can be employed.

In another aspect, the invention provides a pharmaceutical compositioncomprising an antigen binding protein of the present invention or afunctional fragment thereof and a pharmaceutically acceptable carrierfor treatment or prophylaxis of atopic diseases/disorders and chronicinflammatory diseases/disorders, for example, asthma, such as allergicasthma, particularly severe asthma (that is asthma that is unresponsiveto current treatment, including systemically administeredcorticosteroids; see Busse W W et al, J. Allergy Clin. Immunol 2000,106: 1033-1042), “difficult” asthma (defined as the asthmatic phenotypecharacterised by failure to achieve control despite maximallyrecommended doses of prescribed inhaled steroids, see Barnes P J (1998),Eur Respir J 12:1208-1218), “brittle” asthma (defines a subgroup ofpatients with severe, unstable asthma who maintain a wide peakexpiratory flow (PEF) variability despite high doses of inhaledsteroids, see Ayres J G et al (1998) Thorax 58:315-321), nocturnalasthma, premenstrual asthma, steroid resistant asthma (see Woodcock A J(1993) Eur Respir J 6:743-747), steroid dependent asthma (defined asasthma that can be controlled only with high doses of oral steroids),aspirin induced asthma, adult-onset asthma, paediatric asthma.Antibodies of the invention maybe used to prevent, reduce the frequencyof, or mitigate the effects of acute, asthmatic episodes (statusasthmaticus). Antibodies of the invention may also be used to reduce thedosing required (either in terms of amount administered or frequency ofdosing) of other medicaments used in the treatment of asthma. Forexample, antibodies of the invention may be used to reduce the dosingrequired for steroid treatment of asthma such as corticosteroidtreatment (“steroid sparing”). Other diseases or disorders that may betreated with antibodies of the invention include atopic dermatitis,allergic rhinitis, Crohn's disease, chronic obstructive pulmonarydisease (COPD), eosinophilic esophagitis, fibrotic diseases or disorderssuch as idiopathic pulmonary fibrosis, progressive systemic sclerosis(scleroderma), hepatic fibrosis, hepatic granulomas, schistosomiasis,leishmaniasis, and diseases of cell cycle regulation, e.g. Hodgkinsdisease, B cell chronic lymphocytic leukaemia. In one embodiment thedisorder is severe asthma. In a further embodiment the disorder is afibrotic disorder such as IPF.

In a yet further aspect, the invention provides a pharmaceuticalcomposition comprising an antigen binding protein of the presentinvention and a pharmaceutically acceptable carrier for treating atopicdiseases/disorders and chronic inflammatory diseases/disorders, forexample, asthma, such as allergic asthma, particularly severe asthma(that is asthma that is unresponsive to current treatment, includingsystemically administered corticosteroids; see Busse W W et al, J.Allergy Clin. Immunol 2000, 106: 1033-1042), “difficult” asthma (definedas the asthmatic phenotype characterised by failure to achieve controldespite maximally recommended doses of prescribed inhaled steroids, seeBarnes P J (1998), Eur Respir J 12:1208-1218), “brittle” asthma (definesa subgroup of patients with severe, unstable asthma who maintain a widepeak expiratory flow (PEF) variability despite high doses of inhaledsteroids, see Ayres J G et al (1998) Thorax 58:315-321), nocturnalasthma, premenstrual asthma, steroid resistant asthma (see Woodcock A J(1993) Eur Respir J 6:743-747), steroid dependent asthma (defined asasthma that can be controlled only with high doses of oral steroids),aspirin induced asthma, adult-onset asthma, paediatric asthma.Antibodies of the invention maybe used to prevent, reduce the frequencyof, or mitigate the effects of acute, asthmatic episodes (statusasthmaticus). Antibodies of the invention may also be used to reduce thedosing required (either in terms of amount administered or frequency ofdosing) of other medicaments used in the treatment of asthma. Forexample, antibodies of the invention may be used to reduce the dosingrequired for steroid treatment of asthma such as corticosteroidtreatment (“steroid sparing”). Other diseases or disorders that may betreated with antibodies of the invention include atopic dermatitis,allergic rhinitis, Crohn's disease, chronic obstructive pulmonarydisease (COPD), eosinophilic esophagitis, fibrotic diseases or disorderssuch as idiopathic pulmonary fibrosis, progressive systemic sclerosis(scleroderma), hepatic fibrosis, hepatic granulomas, schistosomiasis,leishmaniasis, and diseases of cell cycle regulation, e.g. Hodgkinsdisease, B cell chronic lymphocytic leukaemia. In one embodiment thedisorder is severe asthma. In a further embodiment the disorder is afibrotic disorder such as IPF.

It will be understood that the sequences described herein (SEQ ID NO: 26to SEQ ID NO: 55 and SEQ NO:62 to SEQ ID NO: 146) include sequenceswhich are substantially identical, for example sequences which are atleast 90% identical, for example which are at least 91%, or at least92%, or at least 93%, or at least 94% or at least 95%, or at least 96%,or at least 97% or at least 98%, or at least 99% identical to thesequences described herein.

For nucleic acids, the term “substantial identity” indicates that twonucleic acids, or designated sequences thereof, when optimally alignedand compared, are identical, with appropriate nucleotide insertions ordeletions, in at least about 80% of the nucleotides, at least about 90%to about 95%, or at least about 98% to about 99.5% of the nucleotides.Alternatively, substantial identity exists when the segments willhybridize under selective hybridization conditions, to the complement ofthe strand.

For nucleotide and amino acid sequences, the term “identical” indicatesthe degree of identity between two nucleic acid or amino acid sequenceswhen optimally aligned and compared with appropriate insertions ordeletions. Alternatively, substantial identity exists when the DNAsegments will hybridize under selective hybridization conditions, to thecomplement of the strand.

The percent identity between two sequences is a function of the numberof identical positions shared by the sequences (i.e., % identity=# ofidentical positions/total # of positions times 100), taking into accountthe number of gaps, and the length of each gap, which need to beintroduced for optimal alignment of the two sequences. The comparison ofsequences and determination of percent identity between two sequencescan be accomplished using a mathematical algorithm, as described in thenon-limiting examples below.

The percent identity between two nucleotide sequences can be determinedusing the GAP program in the GCG software package, using a NWSgapdna.CMPmatrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of1, 2, 3, 4, 5, or 6. The percent identity between two nucleotide oramino acid sequences can also be determined using the algorithm of E.Meyers and W. Miller (Comput. Appl. Biosci., 4:11-17 (1988)) which hasbeen incorporated into the ALIGN program (version 2.0), using a PAM120weight residue table, a gap length penalty of 12 and a gap penalty of 4.In addition, the percent identity between two amino acid sequences canbe determined using the Needleman and Wunsch (J. Mol. Biol. 48:444-453(1970)) algorithm which has been incorporated into the GAP program inthe GCG software package, using either a Blossum 62 matrix or a PAM250matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a lengthweight of 1, 2, 3, 4, 5, or 6.

By way of example, a polynucleotide sequence of the present inventionmay be identical to the reference sequence of SEQ ID NO: 25, that is be100% identical, or it may include up to a certain integer number ofnucleotide alterations as compared to the reference sequence. Suchalterations are selected from the group consisting of at least onenucleotide deletion, substitution, including transition andtransversion, or insertion, and wherein said alterations may occur atthe 5′ or 3′ terminal positions of the reference nucleotide sequence oranywhere between those terminal positions, interspersed eitherindividually among the nucleotides in the reference sequence or in oneor more contiguous groups within the reference sequence. The number ofnucleotide alterations is determined by multiplying the total number ofnucleotides in SEQ ID NO: 25 by the numerical percent of the respectivepercent identity (divided by 100) and subtracting that product from saidtotal number of nucleotides in SEQ ID NO: 23, or:

nn≦xn−(xn·y),

wherein nn is the number of nucleotide alterations, xn is the totalnumber of nucleotides in SEQ ID NO: 25, and y is 0.50 for 50%, 0.60 for60%, 0.70 for 70%, 0.80 for 80%, 0.85 for 85%, 0.90 for 90%, 0.95 for95%, 0.97 for 97% or 1.00 for 100%, and wherein any non-integer productof xn and y is rounded down to the nearest integer prior to subtractingit from xn. Alterations of the polynucleotide sequence of SEQ ID NO: 25may create nonsense, missense or frameshift mutations in this codingsequence and thereby alter the polypeptide encoded by the polynucleotidefollowing such alterations.

Similarly, in another example, a polypeptide sequence of the presentinvention may be identical to the reference sequence encoded by SEQ IDNO: 24, that is be 100% identical, or it may include up to a certaininteger number of amino acid alterations as compared to the referencesequence such that the % identity is less than 100%. Such alterationsare selected from the group consisting of at least one amino aciddeletion, substitution, including conservative and non-conservativesubstitution, or insertion, and wherein said alterations may occur atthe amino- or carboxy-terminal positions of the reference polypeptidesequence or anywhere between those terminal positions, interspersedeither individually among the amino acids in the reference sequence orin one or more contiguous groups within the reference sequence. Thenumber of amino acid alterations for a given % identity is determined bymultiplying the total number of amino acids in the polypeptide sequenceencoded by SEQ ID NO: 24 by the numerical percent of the respectivepercent identity (divided by 100) and then subtracting that product fromsaid total number of amino acids in the polypeptide sequence encoded bySEQ ID NO: 24, or:

na≦xa−(xa·y),

wherein na is the number of amino acid alterations, xa is the totalnumber of amino acids in the polypeptide sequence encoded by SEQ ID NO:24, and y is, for instance 0.70 for 70%, 0.80 for 80%, 0.85 for 85%etc., and wherein any non-integer product of xa and y is rounded down tothe nearest integer prior to subtracting it from xa.

The following examples illustrate but do not limit the invention.

EXAMPLES Example 1 Construction of Recombinant Anti-IL-13 Antibodies

Original murine mAbs were produced by immunisation of mice withrecombinant human IL-13. Spleens from responder animals were harvestedand fused to myeloma cells to generate hybridomas. The hybridomasupernatant material was screened for binding. Hybridomas of interestwere monocloned using standard techniques. The resulting murine antibody(6A1) comprises the variable regions shown in SEQ ID NO:58 and SEQ IDNO:59. Further details of this murine antibody and a humanised versionof this antibody A1L1 (SEQ ID NO: 22 and 24) are described inWO2006/003407 which is herein incorporated by reference. The anti-IL-13mAb antibody A1L1 was used in several of the following examples as acomparator antibody.

A number of variants of the humanised antibody comprising the heavychain set out in SEQ ID NO: 22 were produced. These all differed in theCDRH3 region of the antibody (SEQ ID NO: 3).

The base DNA expression constructs for the antibodies of the presentinvention, SEQ ID NO: 23 (heavy chain) and SEQ ID NO:25 (light chain)were prepared de novo by build-up of overlapping oligonucleotidesincluding restriction sites for cloning into Rld and Rln mammalianexpression vectors as well as a human signal sequence. Hind III and SpeI restriction sites were introduced to frame the V_(H) domain containingthe signal sequence (SEQ ID NO:56) for cloning into Rld containing thehuman yl constant region. Hind III and BsiWl restriction sites wereintroduced to frame the V_(L) domain containing the signal sequence (SEQID NO: 56) for cloning into Rln containing the human kappa constantregion. Alternative constructs were produced using pTT vectors whichalso included human constant regions. Where appropriate, site-directedmutagenesis (SDM) was used to generate different humanised constructs.

Example 2 Antibody Expression in HEK 293 6E Cells

pTT plasmids encoding the heavy and light chains respectively weretransiently co-transfected into HEK 293 6E cells and expressed at smallscale to produce antibody. Antibodies were assessed directly from thetissue culture supernatant. Other antibodies were purified usingimmobilised Protein A columns and quantified by reading absorbance at280 nm and where indicated, the purified antibody material was assessedin the assays described in the examples set out below.

Where we refer to the antibodies by code (i.e. A1Y100BTrpL1) we arereferring to the mAb generated by co-transfection and expression of thenoted first and second plasmid, for example ‘A1Y100BTrpL1’ relates to amAb generated by co-transfection of a plasmid containing the A1Y100BTrpsequence and a plasmid containing the L1 sequence in a suitable cellline.

Example 3 Biacore Analysis of Anti IL-13 Humanised mAbs Kinetic Analysis

The initial screen of CDRH3 mutants was carried out on the ProteOn XPR36(Biorad). The method was as follows, antihuman IgG (Biacore BR-1008-39)was immobilised on a GLM chip by primary amine coupling, CDRH3 mutantantibodies were then captured on this surface and IL13 passed over at256, 64, 16, 4, 1 nM with a 0 nM injection (i.e. buffer alone) used todouble reference. 3M MgCl₂ was used to regenerate the capture surface,removing the bound CDRH3 mutant antibodies ready for another cycle ofcapture and analyte injection. The data was fitted to the 1:1 modelusing the software inherent to the machine. All work was carried outusing antibodies directly from tissue culture supernatants except forthe parental antibody which was purified material.

The screen identified several antibodies that appeared to have betterkinetic profile than the parental molecule, these same samples were thenanalysed on the Biacore T100 to confirm the results, using a similarmethod, in that the same antihuman IgG capture antibody was immobilisedon a CM5 chip by primary amine coupling, IL13 was passed over thesurface at 256, 64, 16, 4, 1 and 0.25 nM with a 0 nM used for doublereferencing, regeneration was with 3M MgCl₂ and the data was fitted tothe 1:1 model inherent to the T100. Table 1 details the overallaffinities (equilibrium dissociation constant K_(D)) for the selectedconstructs from the ProteOn screen and the T100 run

TABLE 1 Molecule ProteOn KD (pM) Biacore KD (pM) A1S95TrpL1 NotAnalysable 216 A1I96ValL1 395 696 A1Y97PheL1 683 618 A1D98GluL1 873 779A1H100A AlaL1 Not Analysable 172 A1H100A GluL1 110 195 A1H100A GlnL1 195278 A1H100A ArgL1 256 307 A1H100A SerL1 131 174 A1H100A ThrL1 158 211A1H100A ValL1 112 152 A1Y100B AlaL1 75 83 A1Y100B IleL1 75 73 A1Y100BTrpL1 68 95 A1L1 ~450-600* 366 *A1L1 ran several times during theProteOn screen so range of values obtained

The data highlighted that several mutations at the Y100B residueappeared to improve the overall affinity. In light of this, mutations atthis residue that were not present in the initial screen were testedusing the ProteOn using the same method as described earlier and againusing antibodies direct from tissue culture supernatants. Of themutations tested A1Y100B ValL1, appeared to improve the overall affinity(equilibrium dissociation constant K_(D)) with a value of 0.166 nMobtained compared to a parental value of 0.390-0.460 nM. When Y100B Valwas tested on the Biacore T100 using the same methodology as describedearlier, the equilibrium dissociation constant K_(D) was measured at0.025 nM compared to a parental value of 0.346 nM.

In light of the work so far being carried out using antibody mutantsdirect from tissue culture supernatants, purified antibody was producedfor A1Y100BL1 mutants Ala, Ile and Trp. These were run on the BiacoreT100, using the same method as before and including the A1Y100B ValL1mutant which was not purified at this stage. Table 2 shows the dataobtained from this experiment.

TABLE 2 Molecule ka (M⁻¹s⁻¹) kd (s⁻¹) KD (nM) A1Y100B ValL1 1.018E+63.455E−5 0.034 (supernatant) A1Y100B AlaL1 9.599E+5 3.004E−5 0.031A1Y100B IleL1 1.149E+6 5.584E−5 0.049 A1Y100B TrpL1 2.572E+6 1.627E−40.063 A1L1 1.267E+6 4.560E−4 0.360

The experiment confirmed that the mutants did improve the bindingaffinity to IL13 compared to the parental molecule.

Given that the purified A1Y100BL1 mutants gave better binding affinitiesthan those obtained from tissue culture supernatants, A1Y100B ValL1 waspurified and run along side the other purified A1Y100BL1 mutants thatalso improved affinity using the Biacore T100 machine using the methoddescribed earlier. Table 3, shows the data obtained from thisexperiment. This experiment was in good agreement with the data in Table2 and confirmed the improvement of affinity for the Y100B mutations.

TABLE 3 Molecule ka (M⁻¹s⁻¹) kd (s⁻¹) KD (nM) A1Y100B IleL1 (pur)9.886E+5 4.214E−5 0.043 A1Y100B ValL1 (pur) 7.757E+5 2.123E−5 0.027A1Y100B AlaL1 (pur) 8.096E+5 2.583E−5 0.032 A1Y100B TrpL1 (pur) 2.385E+61.253E−4 0.053 A1L1(pur) 1.128E+6 3.677E−4 0.326

Example 4 Neutralisation of E. Coli-Expressed Recombinant Human IL-13 ina TF-1 Cell Proliferation Bioassay

TF-1 cells proliferate in response to a number of different cytokinesincluding human IL-13. The proliferative response of these cells forIL-13 can therefore be used to measure the bioactivity of IL-13 andsubsequently an assay has been developed to determine the IL-13neutralisation potency (inhibition of IL-13 bioactivity) of mAbs.

The assay was performed in sterile 96-well tissue culture plates understerile conditions and all test wells were performed in triplicate. 14ng/ml recombinant E. Coli-expressed human IL-13 was pre-incubated withvarious dilutions of mAbs (usually from 93.4 nM titrated in 3-folddilutions to 0.014 nM) for 1 hour at 37° C. An antibody of irrelevantspecificity was similarly titrated as a negative control. These sampleswere then added to 50 μl of TF-1 cells (at a concentration of 2×10⁵cells per ml) in a sterile 96-well tissue culture plate. Thus the final100 μl assay volume contained various dilutions of mAbs (at a finalconcentration of 46.7 nM titrated in 3-fold dilutions to 0.007 nM),recombinant E. Coli-expressed human IL-13 (at a final concentration of 7ng/ml) and TF-1 cells (at a final concentration of 1×10⁵ cells per ml).The assay plate was incubated at 37° C. for approximately 3 days in ahumidified CO₂ incubator. The amount of cell proliferation was thendetermined using the ‘CellTitre 96® Non-Radioactive Cell ProliferationAssay’ from Promega (catalogue number G4100), as described in themanufacturers instructions. The absorbance of the samples in the 96-wellplate was read in a plate reader at 570 nm.

The capacity of the mAbs to neutralise recombinant E. Coli-expressedhuman IL-13 bioactivity was expressed as that concentration of the mAbrequired to neutralise the bioactivity of the defined amount of humanIL-13 (7 ng/ml) by 50% (=ND₅₀). The lower the concentration of the mAbrequired, the more potent the neutralisation capacity. The ND₅₀ dataprovided herein (Table 4) were calculated using Robosage in MicrosoftExcel. Graphical representation of the data can be seen in FIG. 1.

TABLE 4 Molecule ND₅₀ (nM) Standard Error (nM) A1L1 9.77 34.82A1Y100BAla L1 0.92 0.08 A1Y100BIle L1 1.10 0.21 A1Y100BTrpL1 1.25 0.34

Example 5 Construction and Expression of mAb-dAbs Comprising the CDRH3Variant Anti-IL-13 mAb

Using standard molecular biology techniques, genes encoding each of thesequences for the variable heavy regions of the CDRH3 variants of the A1antibodies were transferred from existing constructs to an expressionvector containing the hIgG1 constant region fused to an anti-human IL-4domain antibody (DOM9-1,2-210) via a TVAAPS or ASTKGPS linker at thec-terminus of the hIgG1 constant region. Details of the heavy chainsconstructed are listed in Table 5.

TABLE 5 Molecule Protein DNA number Name Description Seq ID Seq IDBPC1624 A1Y100BAla H- H Chain = A1Y100B Ala H 62 63 TVAAPS-210 L1chain-TVAAPS linker-DOM9- 112-210 dAb L chain = L1 24 25 BPC1625A1Y100BIle H- H chain = A1Y100BIle H chain- 64 65 TVAAPS-210 L1TVAAPS linker-DOM9-112-210 dAb L chain = L1 24 25 BPC1626 A1Y100BTrp H-H chain = A1Y100BTrp H chain- 66 67 TVAAPS-210 L1TVAAPS linker-DOM9-112-210 dAb L chain = L1 24 25 BPC1627 A1Y100BVal H-H chain = A1Y100BVal H chain- 68 69 TVAAPS-210 L1TVAAPS linker-DOM9-112-210 dAb L chain = L1 24 25 BPC1628 A1Y100BAla H-H Chain = A1Y100B Ala H 70 71 ASTKGPS-210 L1 chain-ASTKGPS linker-DOM9-112-210 dAb L chain = L1 24 25 BPC1629 A1 Y100BIle H- H chain =A1Y100BIle H chain- 72 73 ASTKGPS-210 L1 ASTKGPS linker-DOM9-112-210 dAb L chain = L1 24 25 BPC1630 A1Y100BTrp H- H chain =A1Y100BTrp H chain- 74 75 ASTKGPS-210 L1 ASTKGPS linker-DOM9-112-210 dAb L chain = L1 24 25 BPC1631 A1Y100BVal H- H chain =A1Y100BVal H chain- 76 77 ASTKGPS-210 L1 ASTKGPS linker-DOM9-112-210 dAb L chain = L1 24 25

BPC1624, BPC1625, BPC1626 and BPC1627 were expressed in HEK293 cells.Briefly, 250 ml of HEK293 cells at 1.5×10⁶ cells/ml were co-transfectedwith heavy and light chain expression plasmids previously incubated with293fectin reagent (Invitrogen #51-0031). These were placed in a shakingincubator at 37° C., 5% CO₂, and 95% relative humidity. After 24 hoursTryptone feeding media was added and the cells grown for a further 5days. Supernatant was harvested by centrifugation and filter sterilised.The expressed molecules were purified by affinity chromatography usingimmobilised Protein A columns and the concentration was determined bymeasuring the absorbance at 280 nm. The level of aggregated protein inthe purified samples was determined by size exclusion chromatography.The yield of purified protein and levels of aggregation are shown inTable 5b.

TABLE 5b BPC Name Yield % aggregate BPC1624 586Y100BA H-TVAAPS-210 0.81mg 3.6% BPC1625 586Y100BI H-TVAAPS-210 0.944 mg  6.7% BPC1626 586Y100BWH-TVAAPS-210 1.14 mg 5.5% BPC1627 586Y100BV H-TVAAPS-210 1.26 mg 8.4%

Example 6 Neutralisation Activity Data for mAb-dAbs Comprising the CDRH3Variant Anti-IL-13 mAb

mAb-dAbs comprising the CDRH3 variant anti-IL-13 mAb were tested forneutralisation of E. Coli-expressed recombinant human IL-13 in a TF-1cell proliferation bioassay

The assay was performed in sterile 96-well tissue culture plates understerile conditions and all test wells were performed in triplicate.Approximately 20 ng/ml recombinant E. Coli-expressed human IL-13 waspre-incubated with various dilutions of mAbdAbs (usually from 50 nMtitrated in 3-fold dilutions to 0.02 nM) (those mAbdAbs made in HEKcells and purified as described in example 5) in a total volume of 50 μlfor 1 hour at 37° C. An antibody of irrelevant specificity was similarlytitrated as a negative control (data not shown). These samples were thenadded to 50 μl of TF-1 cells (at a concentration of 2×10⁵ cells per ml)in a sterile 96-well tissue culture plate. Thus the final 100 μl assayvolume contained various dilutions of mAbdAbs (at a final concentrationof 25 nM titrated in 3-fold dilutions to 0.01 nM), recombinant E.Coli-expressed human IL-13 (at a final concentration of 10 ng/ml) andTF-1 cells (at a final concentration of 1×10⁵ cells per ml). The assayplate was incubated at 37° C. for approximately 3 days in a humidifiedCO₂ incubator. The amount of cell proliferation was then determinedusing the ‘CellTitre 96® Non-Radioactive Cell Proliferation Assay’ fromPromega (catalogue number G4100), as described in the manufacturer'sinstructions. The absorbance of the samples in the 96-well plate wasread in a plate reader at 570 nm.

The capacity of the mAbdAbs to neutralise human IL-13 bioactivity wasexpressed as that concentration of the mAb-dAb required to neutralisethe bioactivity of the defined amount of human IL-13 (10 ng/ml) by 50%(=ND₅₀). The lower the concentration of the mAbdAb required, the morepotent the neutralisation capacity. The ND₅₀ data provided herein (Table6) were calculated using GraphPad Prism. These data are representedgraphically in FIG. 2.

TABLE 6 Antibody ID Description ND₅₀ human IL-13 BPC1624 A1Y100BAlaH-TVAAPS-210L1 0.553 nM BPC1625 A1Y100BIle H-TVAAPS-210L1 0.542 nMBPC1626 A1Y100BTrp H-TVAAPS-210L1 0.681 nM BPC1627 A1Y100BValH-TVAAPS-210L1 0.615 nM A1L1 Anti IL-13 antibody 2.524 nM

Example 7 Expression of mAb-dAbs Comprising the CDRH3 Variant Anti-IL-13mAb in the CHOE1a Expression System

Molecules BPC 1624 to 1631 as shown in Table 5 were also expressed inCHOE1a cells. DNA vectors encoding the heavy and light chains wereco-electroporated into suspension CHO cells. Cells were passaged inshake flasks in MR1 basal selective medium at 37° C., 5% CO₂, 130 rpmuntil cell viability and cell counts improved. CHO cells were theninoculated into MR1 basal x2 selective medium and incubated for 8 to 12days at 34° C., 5% CO₂, 130 rpm. The cells were pelleted bycentrifugation and the supernatant sterile filtered.

Expressed material was purified by affinity chromatography usingimmobilised protein A columns and the yield determined by measurement ofabsorbance at 280 nm. The level of aggregates was determined by sizeexclusion chromatography. Aggregates were removed by preparative sizeexclusion chromatography and the yield re-assessed. Table 7 lists theyields and levels of aggregate obtained from this expression system.

TABLE 7 Yield Aggregates (mgs) (%) (post (post protein protein FinalFinal Expression A pre A pre Yield Aggregates Molecule Volume (ml)clean-up) clean-up) (mgs) (%) BPC1628 850 80.70 17.1% 39.50 2.6% BPC1629850 77.90 20.3% 35.25 2.8% BPC1630 850 69.30 15.7% 38.00 3.3% BPC1631850 61.47 20.3% 30.00 2.3% BPC1624 850 88.65 15.2% 45.00 2.6% BPC1625850 77.49 16.2% 37.49 2.3% BPC1626 850 65.16 12.6% 30.80 3.1% BPC1627850 73.26 15.2% 34.44 1.9%

Example 8 Stoichiometry Assessment of Antigen Binding Proteins (UsingBiacore™)

This example is prophetic. It provides guidance for carrying out anadditional assay in which the antigen binding proteins of the inventioncan be tested,

Anti-human IgG is immobilised onto a CM5 biosensor chip by primary aminecoupling. Antigen binding proteins are captured onto this surface afterwhich a single concentration of IL-13 or IL-4 or IL-5 is passed over,this concentration is enough to saturate the binding surface and thebinding signal observed reached full R-max. Stoichiometries are thencalculated using the given formula:

Stoich=Rmax*Mw(ligand)/Mw(analyte)*R(ligand immobilised or captured)

Where the stoichiometries are calculated for more than one analytebinding at the same time, the different antigens are passed oversequentially at the saturating antigen concentration and thestoichometries calculated as above. The work can be carried out on theBiacore 3000, at 25° C. using HBS-EP running buffer.

Example 9 Dose Prediction of Improved Humanised Variant MAbs

An antibody-ligand binding PK-PD model was developed in order to rankthe different monoclonal antibody (mAb) candidates based on bindingaffinity and predicted potential therapeutic dose in human.

The predicted potential therapeutic dose in human was defined for thispurpose as the dose providing 90% inhibition of the target IL-13 in thelung (site of action) at steady-state following monthly intravenousadministration of the mAbs for 1 h. The molecular weight of eachmolecule was assumed to be the same and equal to the standard molecularweight of a mAb i.e. 150 kDa. In addition, in the absence of animal orhuman pharmacokinetics data for the different candidates, the humanpharmacokinetics of the A1L1 antibody was inferred to all thecandidates.

The same antibody-ligand binding PK-PD model is used for each mAb aswell as the same assumptions regarding the target concentration, thetarget turnover, the target tissue:plasma ratio and the mAb tissuepenetration. The ranking provided by the model is therefore solely basedon the binding affinity of the molecules, the only parameter differing.In such conditions, the potential therapeutic dose in human for the 4candidates A1Y100BIleL1, A1Y100BValL1, A1Y100BAlaL1 and A1Y100BTrpL1 ispredicted to provide a substantial improvement above the predictedpotential therapeutic dose in human for A1L1.

Example 10 Anti-IL13/IL4 mAbdAbs with Variant IL-4 dAbs 10.1Construction and Expression

The anti IL-4 dAb (DOM9-155-154, SEQ ID NO: 80), was investigated foraggregation-prone residues using an aggregation prediction algorithm.The leucine residue, at Kabat position 89 was identified as a keyresidue for promotion of aggregation.

In order to reduce the aggregation potential of mAbdAbs containing thisdAb, this amino acid residue was substituted for other amino acids togenerate a number of mAb-dAb variants. Expression constructs weregenerated by site directed mutagenesis using the DNA expression vectorcoding for the heavy chain of an existing mAbdAb construct. The proteinsequences for the resulting new mAbdAb heavy chains comprising themutated dAb sequences are given in SEQ ID NOs 117-134.

Other heavy chain sequences incorporating another mutation at position89 are SEQ ID NOs: 96-106. These are described in detail in Example 11.

Table 8 provide a list of the molecules expressed.

TABLE 8 Protein Alternative SEQ ID Identifier names LinkerMolecule description NO: BPC1090 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 117 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89G) 155-154 (89G)L chain: Anti-human IL-13 mAb light 24 chain BPC1091 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 118 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89S) 155-154 (89S)L chain: Anti-human IL-13 mAb light 24 chain BPC1092 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 119 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89H) 155-154 (89H)L chain: Anti-human IL-13 mAb light 24 chain BPC1093 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 147 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89M) 155-154 (89M)L chain: Anti-human IL-13 mAb light 24 chain BPC1094 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 121 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89A) 155-154 (89A)L chain: Anti-human IL-13 mAb light 24 chain BPC1095 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 122 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89T) 155-154 (89T)L chain: Anti-human IL-13 mAb light 24 chain BPC1108 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 123 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89C) 155-154 (89C)L chain: Anti-human IL-13 mAb light 24 chain BPC1109 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 124 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89R) 155-154 (89R)L chain: Anti-human IL-13 mAb light 24 chain BPC1110 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 125 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89W) 155-154 (89W)L chain: Anti-human IL-13 mAb light 24 chain BPC1111 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 126 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89E) 155-154 (89E)L chain: Anti-human IL-13 mAb light 24 chain BPC1112 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 127 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89K) 155-154 (89K)L chain: Anti-human IL-13 mAb light 24 chain BPC1113 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 128 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89D) 155-154 (89D)L chain: Anti-human IL-13 mAb light 24 chain BPC1114 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 129 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89N) 155-154 (89N)L chain: Anti-human IL-13 mAb light 24 chain BPC1115 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 130 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89Y) 155-154 (89Y)L chain: Anti-human IL-13 mAb light 24 chain BPC1116 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 131 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89V) 155-154 (89V)L chain: Anti-human IL-13 mAb light 24 chain BPC1117 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 132 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89I) 155-154 (89I)L chain: Anti-human IL-13 mAb light 24 chain BPC1118 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 133 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89F) 155-154 (89F)L chain: Anti-human IL-13 mAb light 24 chain BPC1119 829H- (TVAAPS)₂GSH chain: Anti-human IL-13 mAb heavy 134 (TVAAPS)₂GS-154chain Y100B_V-(TVAAPS)₂GS-DOM9- (89P) 155-154 (89P)L chain: Anti-human IL-13 mAb light 24 chain

10.2 Molecule Expression in HEK 293 6E Cells

Plasmids encoding the heavy and light chains respectively weretransiently co-transfected into HEK 293 6E cells and expressed at smallscale to produce antibody molecules. A tryptone feed was added to eachcell culture up to 24 hours after transfection and the cells wereharvested after 3 days.

Antibody molecules were assessed directly from the tissue culturesupernatant and quantified using the Gyrolab workstation.

Gyrolab Worstation Method for Quantification of Antibody Molecules inCell Supernatant

Antibodies produced from small scale transient HEK 2936E transfections(0.75-2.0 ml) were quantified from tissue culture supernatants by aquantitative immunoassay using a Gyrolab Bioaffy Workstation (Gyros).Antibody was captured via the Fc region using a biotinylated anti-IgGAffibody molecule (Abcam) immobilised onto streptavidin-coated particleson a compact disc (CD) microlaboratory (Gyros). The Affibody reagent wasvortexed briefly and diluted with PBS-Tween 20 (0.01% v/v) to a finalworking concentration of 0.1 mg/ml. Antibody was then detected by anALEXA 647 labelled Fab2 anti-human IgG kappa light chain molecule usinglaser-induced fluorescence. The ALEXA 647 labelled detection reagent wasprepared by vortexing briefly and by centrifugation at 13000 rpm for 4minutes. The labelled Fab2 detection reagent was added to unlabelledFab2 which were diluted to final concentrations of 75 nM and 1.5 μMrespectively using Rexcip F Detection reagent diluant (Gyros). Theantibody quantification range was between 0.244-250 μg/ml relative to ananti-CD23 monoclonal antibody standard curve. The anti-CD23 (1 mg/ml)standard curve was generated by serial dilution of the antibody withtissue culture media (Freestyle 293 Expression Media, Pluronic F68 andGeneticin, Invitrogen).

In some instances, the antibody molecules were purified usingimmobilised Protein A columns and quantified by reading absorbance at280 nm and where indicated, the purified antibody molecule was assessedin the assays described in the examples set out below.

10.3 IL-4 Binding ELISA

These mAbdAbs were tested for binding to IL-4 in a direct binding ELISAusing the following method.

96-well high binding plates were coated with 5 μg/ml human IL-4 (made atGSK) in NaHCO₃ and stored overnight at 4° C. The plates were washedtwice with Tris-Buffered Saline with 0.05% of Tween-20 (TBST). 100 μL ofblocking solution (1% BSA in TBST buffer) was added in each well and theplates were incubated for at least one hour at room temperature. ThemAbdAbs were successively diluted across the plates in blockingsolution. After one hour incubation, the plates were washed three times.Goat anti-human kappa light chain specific peroxidase conjugatedantibody (Sigma A7164) was diluted in blocking solution to 1 μg/mL and50 μL was added to each well. The plates were incubated for one hour.After another three washing steps, 50 μl of OPD (o-phenylenediaminedihydrochloride) SigmaFast substrate solution was added to each well andthe reaction was stopped after about 5 minutes by addition of 25 μL of3M sulphuric acid. Absorbance was read at 490 nm using the VersaMaxTunable Microplate Reader (Molecular Devices) using a basic endpointprotocol.

The experiment was carried out using mAbdAbs directly from tissueculture supernatants which had been quantified using the gyrolabplatform except for the positive control (anti-IL-4 mAb) and theanti-IL13 negative control mAb, which were purified material. These dataare shown in FIG. 17.

The result of the ELISA shows that most of these transiently expressedanti-IL13 mAb-anti-IL4 dAbs bound IL-4, but some variation in IL-4binding activity was observed. The purified positive control anti-IL-4mAb, also showed binding to IL-4, whereas the purified negative controlmAb showed no binding to human IL-4.

10.4 Analysis of Levels of Aggregate in mAbdab Expressions

pTT plasmids encoding the heavy and light chains of BPC1090, BPC1091,BPC1092, BPC1093, BPC1094 and BPC1095 were transiently co-transfectedinto HEK 293 6E cells and expressed using 293fectin (Invitrogen,12347019) at slightly larger scale (between 200 and 600 ml) than themAbdabs described above in Example 10.2. The BPC1111 and BPC1085 wereindependently transiently expressed in HEK293 6E cells using the samemethodology. The plasmids used for the above transfections weregenerated using the EndoFree Plasmid Maxi Kit (Qiagen, 12362).

A tryptone feed was added to each of the cell cultures after 24 hoursand the cells were harvested after 72 hours. The antibodies werepurified using a Protein A column, quantified by reading absorbance at280 nm and analyzed by size exclusion chromatography (SEC).

These mAbdAbs were compared with BPC2223, anti-IL13 mAb (829) andanti-II-13 mAb (586) which had been independently expressed.

Both antibodies (586 with the original CDRH3 and 829 with the mutatedCDRH3) showed low levels of aggregation, as did the mAbdAbs comprisingthe mutated dAb (BPC1090, BPC 1091, BPC 1093, BPC1094 and BPC1095).BPC2223 which comprised the original dAb i.e. where position 89 was notmutated had higher levels of aggregation, as did BPC1092 which had anL89H mutation. Representative aggregation data is shown in Table 8b.

TABLE 8b Molecule % aggregates number Molecule description (SEC) BPC1085829H-GS(TVAAPSGS)₂-154 (89Q) ~1.6 BPC 1090 829H-(TVAAPS)₂GS-154 (89G)~0.7 BPC 1091 829H-(TVAAPS)₂GS-154 (89S) ~1.5 BPC 1092829H-(TVAAPS)₂GS-154 (89H) ~20 BPC 1093 829H-(TVAAPS)₂GS-154 (89M) ~4.3BPC 1094 829H-(TVAAPS)₂GS-154 (89A) ~0.8 BPC 1095 829H-(TVAAPS)₂GS-154(89T) ~2 BPC 1111 829H-(TVAAPS)₂GS-154 (89E)  <5% BPC 2223586H-GS(TVAAPSGS)₂-154 ~25% — 829 (mAb) ~2.1 — 586 (mAb) <1

10.5 BIAcore Analysis

Purified mAbs and mAbdAb constructs were tested in a BIAcore assay todetermine whether the mutation of position 89 had any effect on thebinding of the dAb to IL-4.

Protein A was immobilised on a CM5 chip by primary amine coupling; thissurface was used as a capture surface for the antibody molecules to betested. Recombinant E. coli-expressed Human IL4 was used as analyte at256, 64, 16, 4 and 1, 0.25 and 0.0625 with 0 nM (i.e. buffer alone) usedto double reference the binding curves. Regeneration of the anti-ProteinA surface was achieved using 50 mM NaOH. The assay was run at 25° C.using HBS-EP as running buffer. The data was fitted to 1:1 modelinherent to the Biacore T100 analysis software.

TABLE 8c Molecule KD Number Molecule Description ka (1/Ms) kd (1/s) (nM)BPC1085 829H-GS(TVAAPSGS)₂- 2.38E+07 1.23E−03 0.052 256 BPC2223586H-GS(TVAAPSGS)₂- 9.71E+06 9.34E−05 0.010 154 BPC1090829H-(TVAAPS)₂GS-154 3.00E+06 4.55E−04 0.152 (89G) BPC1091829H-(TVAAPS)₂GS-154 3.96E+06 9.66E−04 0.244 (89S) BPC1092829H-(TVAAPS)₂GS-154 2.98E+06 5.34E−03 1.794 (89H) BPC1093829H-(TVAAPS)₂GS-154 4.19E+06 2.16E−04 0.051 (89M) BPC1094829H-(TVAAPS)₂GS-154 1.91E+06 3.73E−03 1.948 (89A) BPC1095829H-(TVAAPS)₂GS-154 2.50E+06 1.44E−02 5.760 (89T) BPC1111829H-(TVAAPS)₂GS-154 3.37E+06 1.04E−04 0.031 (89E)

Example 11 Construction and Testing of Antigen Binding ProteinsComprising the CDRH3 Variant Anti-IL-13 mAb and a Mutated dAb (BPC1085,BPC1086 & BPC10871 11.1 Construction and Expression

Plasmids encoding heavy chains consisting of an anti-IL-13 mAb and ananti-IL-4 dAb were used as base constructs to generate alternativeplasmid constructs. A two step cloning strategy was required. In step 1,the DNA sequence encoding the VH of the anti-IL13 mAb component of the Hchain was replaced with the DNA sequence encoding the VH of anotherhumanized anti-IL13 antibody (SEQ ID NO:54) by restriction cloning usingHindIII and SpeI. In step 2, the codon encoding the leucine at Kabatposition 89 in the anti-IL4 dAb (DOM9-155-154, SEQ ID NO: 80) componentof the mAbdAb was mutated by site directed mutagenesis to glutamine. Allof the resulting heavy chain DNA sequences generated are given in SEQ IDNOs: 96, 98 and 100. Table 9 provides a list of the moleculesconstructed and expressed.

TABLE 9 Summary of the antibodies constructed and expressed AlternativeProtein Identifier names Linker Molecule description SEQ ID NO: BPC1085829H- GS(TVAAPSGS)₂ H chain: Anti-human IL- 96 GS(TVAAPSGS)₂-13 mAb heavy chain 154 (89Q) or Y100B_V- 829H- GS(TVAAPSGS)₂-DOM9-GS(TVAAPSGS)₂- 155-256 256 L chain: Anti-human IL-13 24 mAb light chainBPC1086 829H- GS(TVAAPSGS)₃ H chain: Anti-human IL- 98 GS(TVAAPSGS)₃-13 mAb heavy chain 154 (89Q) or Y100B_V- 829H- GS(TVAAPSGS)₃-DOM9-GS(TVAAPSGS)₃- 155-256 256 L chain: Anti-human IL-13 24 mAb light chainBPC1087 829H- GS(TVAAPSGS)₄ H chain: Anti-human IL- 100 GS(TVAAPSGS)₄-13 mAb heavy chain 154 (89Q) or Y100B_V- 829H- GS(TVAAPSGS)₄-DOM9-GS(TVAAPSGS)₄- 155-256 256 L chain: Anti-human IL-13 24 mAb light chain

Heavy and light chain expression plasmids encoding BPC1085, BPC1086 andBPC1087 mAbdAbs were co-transfected into HEK 2936E cells using 293fectin(Invitrogen, 12347019). A tryptone feed was added to each of the cellcultures after 24 hours and the cells were harvested after 72 hours. Theantibodies were purified using a Protein A column before being tested inbinding assays.

BPC1085, BPC1086 and BPC1087 mAbdAbs were purified using Protein Aaffinity. 1 ml Protein A columns were used (GE Healthcare) on the AKTAXpress system, columns were equilibrated in PBS (Gibco/Invitrogen) andthe antibodies eluted using Pierce IgG elute. Eluted fractions wereneutralised using 1M Tris (Hydroxymethyl) Aminomethane buffer (ingeneral 5-10% v/v). Eluted antibody fractions were pooled and analysedfor aggregation by size exclusion chromatography and quantified byreading at OD₂₈₀ nm using a spectrophotometer.

These were compared to equivalent mAbdAbs (2222, 2223, 2230 and 2231)which are described in Table 10. These comprise:

-   -   i) a dAb which is identical to that used in BPC1085, BPC1086 and        BPC1087 except for position 89 which is ‘L’ in BPC2222, BPC2223,        BPC2230 & BPC2231 and ‘Q’ in BPC1085, BPC1086 & BPC1087).    -   ii) same linkers    -   iii) an IL-13 mAb sequence which is identical to that used        BPC1085, BPC1086 and BPC1087 except for position 100B which is        ‘Y’ in BPC2222, BPC2223, BPC2230 & BPC2231 and ‘V’ in BPC1085,        BPC1086 & BPC1087).

TABLE 10 Identi- Heavy Light fier mAb Linker dAb chain chain BPC2222A1L1 GS DOM9- 135 24 (TVAAPSGS)₁ 155-154 BPC2223 A1L1 GS DOM9- 136 24(TVAAPSGS)₂ 155-154 BPC2230 A1L1 GS DOM9- 137 24 (TVAAPSGS)₃ 155-154BPC2231 A1L1 GS DOM9- 138 24 (TVAAPSGS)₄ 155-154

BPC2222, 2223, 2230 and 2231 mAbdAbs were purified using Protein Aaffinity. 1 ml Protein A columns were used (GE Healthcare) on the AKTAXpress system, columns were equilibrated in PBS (Gibco/Invitrogen) andthe antibodies eluted using Pierce IgG elute. Eluted fractions wereneutralised using 1M Tris (Hydroxymethyl) Aminomethane buffer (ingeneral 5-10% v/v). Eluted antibody fractions were pooled and analysedfor aggregation by size exclusion chromatography and quantified byreading at OD₂₈₀ nm using a spectrophotometer.

BPC2222, 2223, 2230 and 2231 showed aggregation of between 30-40%, withthe aggregated material eluting before 10 minutes.

Compared to BPC2222, 2223, 2230 and 2231 the constructs BPC1085, 1086and 1087 showed lower levels of aggregation as assessed by sizeexclusion chromatography. The SEC profiles for these molecules are shownin FIGS. 3-9

11.2 IL-4 Binding ELISA

Purified BPC1085, BPC1086 and BPC1087 mAbdAbs were tested for binding toIL-4 in a direct binding ELISA according to the method described inExample 10.3.

These data are shown in FIG. 10. The results of the ELISA confirmed thatthese purified mAbdAbs bound to IL-4. The positive controls anti-IL-4mAb and BPC2231 also showed binding to IL-4 whereas the negative controlmAb (anti IL-13 mAb) showed no binding to IL-4. This indicated in thisELISA the dAb potency increased when the linker length was increasedfrom GS(TVAAPSGS) to GS(TVAAPSGS)₂₋₄.

11.3 Neutralization of IL-4 in a TF-1 Cell Proliferation Bioassay

Purified BPC1085, BPC1086 and BPC1087 mAbdAbs were tested forneutralization of human IL-4 in a TF-1 cell bioassay.

TF-1 cells proliferate in response to a number of different cytokinesincluding human IL-4. The proliferative response of these cells for IL-4can therefore be used to measure the bioactivity of IL-4 andsubsequently an assay has been developed to determine the IL-4neutralisation potency (inhibition of IL-4 bioactivity) of mAbdAbs.

The assay was performed in sterile 96-well tissue culture plates understerile conditions and all test wells were performed in duplicate.Approximately 2.2 ng/ml recombinant E. Coli-expressed human IL-4 waspre-incubated with various dilutions of mAbdAbs (usually from 560 nMtitrated in 3-fold dilutions to 0.009 nM) in a total volume of 120 μlfor 1 hour at 37° C. An antibody of irrelevant specificity was similarlytitrated as a negative control (anti-IL13 mAb). 50 μl of these sampleswere then added to 50 μl of TF-1 cells (at a concentration of 2×10⁵cells per ml) in a sterile 96-well tissue culture plate. Thus the final100 μl assay volume contained various dilutions of mAbdAbs (at a finalconcentration of 270 nM titrated in 3-fold dilutions to 0.005 nM),recombinant E. Coli-expressed human IL-4 (at a final concentration of1.1 ng/ml) and TF-1 cells (at a final concentration of 1×10⁵ cells perml). The assay plate was incubated at 37° C. for approximately 4 days ina humidified CO₂ incubator. The amount of cell proliferation was thendetermined using the ‘CellTitre 96® Non-Radioactive Cell ProliferationAssay’ from Promega (catalogue number G4100), as described in themanufacturers instructions. The absorbance of the samples in the 96-wellplate was read in a plate reader at 570 nm. These data were entered onan Excel spreadsheet, values for duplicate test wells were averaged andthe average background value (no mAb-dAb and no IL-4 test wells) wassubtracted.

The capacity of the mAbdAbs to neutralise recombinant E. Coli-expressedhuman IL-4 bioactivity was expressed as that concentration of themAb-dAb required to neutralise the bioactivity of the defined amount ofhuman IL-4 (1.1 ng/ml) by 50% (=ND₅₀). The lower the concentration ofthe mAbdAb required, the more potent the neutralisation capacity. TheND₅₀ data provided herein (Table 11) were calculated using the Robosagefunction in Excel. These data are represented graphically in FIG. 11.

An anti-IL-4 mAb and DOM9-155-154 (SEQ ID NO: 80) were included aspositive controls for neutralization of human and cynomolgus IL-4 in theTF-1 cell bioassays. Additionally, a dAb with specificity for anirrelevant antigen (dummy dAb) was also included as a negative controlfor neutralization of human or cynomolgus IL-4 in the TF-1 cellbioassays.

These were repeated several times and FIG. 11 shows the results for oneof these experiment s. ND₅₀ values were calculated from the dataset. TheND₅₀ value is the concentration of the mAbdAb or mAb or dAb, which isable to neutralise the bioactivity of IL-4 by 50%. The mean ND₅₀ valueand the number of times tested (n) are shown in Table 11.

TABLE 11 Mean ND₅₀ value & standard Number Molecule deviation (nM) ofrepeats BPC1085 9.21 3 BPC1086 4.32 3 BPC1087 3.59 3 anti-IL-4 mAb 1.952 DOM9-155-154 0.98 2 Negative control dAb did not 2 neutralise

These data confirm that purified BPC1085, BPC1086 and BPC1087 mAbdAbs,neutralized the bioactivity of human and cyno IL-4. Anti-IL-4 mAb andDOM9-155-154 also neutralised the bioactivity of human and cynomolgusIL-4, whereas the negative dAb (dummy dAb) showed no neutralization inthe same bioassay.

All three mAbdAbs show good potency, and there is a clear trend ofincreasing dAb potency with increasing linker length was apparent fromthe neutralisation assays, despite the more crude ELISA not picking upthis difference in potency. A negative control anti-IL-4 mAb) showed noneutralization in the same bioassay.

11.4 Neutralization of Human IL-13 in a TF-1 Cell Proliferation Bioassay

Purified BPC1085, BPC1086 and BPC1087 mAbdAbs were tested forneutralization of human IL-13 in a TF-1 cell bioassay as describedbelow.

TF-1 cells proliferate in response to a number of different cytokinesincluding human IL-13. The proliferative response of these cells forIL-13 can therefore be used to measure the bioactivity of IL-13 andsubsequently an assay has been developed to determine the IL-13neutralisation potency (inhibition of IL-13 bioactivity) of mAbdAbs. Theassay was performed in sterile 96-well tissue culture plates understerile conditions and all test wells were performed in duplicate.Approximately 14 ng/ml recombinant E. Coli-expressed human IL-13 waspre-incubated with various dilutions of mAbdAbs (usually from 560 nMtitrated in 3-fold dilutions to 0.009 nM) in a total volume of 120 μlfor 1 hour at 37° C. An antibody and dAb of irrelevant specificity wassimilarly titrated as negative controls (anti-IL-4 mAb and DOM9-155-154respectively). 50 μl of these samples were then added to 50 μl of TF-1cells (at a concentration of 2×10⁵ cells per ml) in a sterile 96-welltissue culture plate. Thus the final 100 μl assay volume containedvarious dilutions of mAbdAbs (at a final concentration of 270 nMtitrated in 3-fold dilutions to 0.005 nM), recombinant E. Coli-expressedhuman IL-13 (at a final concentration of 7 ng/ml) and TF-1 cells (at afinal concentration of 1×10⁵ cells per ml). The assay plate wasincubated at 37° C. for approximately 4 days in a humidified CO₂incubator. The amount of cell proliferation was then determined usingthe ‘CellTitre 96® Non-Radioactive Cell Proliferation Assay’ fromPromega (catalogue number G4100), as described in the manufacturer'sinstructions. The absorbance of the samples in the 96-well plate wasread in a plate reader at 570 nm. These data were entered on an Excelspreadsheet, values for duplicate test wells were averaged and theaverage background value (no mAb-dAb and no IL-13 test wells) wassubtracted. The capacity of the mAbdAbs to neutralise recombinant E.Coli-expressed human IL-13 bioactivity was expressed as thatconcentration of the mAb-dAb required to neutralise the bioactivity ofthe defined amount of human IL-13 (7 ng/ml) by 50% (=ND₅₀). The lowerthe concentration of the mAbdAb required, the more potent theneutralisation capacity. The ND₅₀ data provided herein (Table 12) werecalculated using the Robosage function in Excel. These data arerepresented graphically in FIG. 12.

An anti IL-13 mAb (SEQ ID NO:22 & 24) was included as a positive controlfor neutralization of human IL-13 in the TF-1 cell bioassays.Additionally, an anti-IL-4 mAb was also included as a negative control.

FIG. 12 shows the result of the TF-1 cell neutralization assay. ND₅₀values were calculated from the dataset. The ND₅₀ value is theconcentration of the mAbdAb or mAb or dAb, which is able to neutralisethe bioactivity of IL-13 by 50%. The mean ND₅₀ value and the number oftimes tested are shown in Table 12.

TABLE 12 Mean ND₅₀ value & standard Number of Molecule deviation (nM)repeats BPC1085 0.88 1 BPC1086 1.01 1 BPC1087 1.14 1 Anti-IL-13 mAb 5.011 anti-IL-4 mAb did not neutralise 1

These data confirm that purified BPC1085, BPC1086 and BPC1087 mAbdAbs,neutralized the bioactivity of recombinant human and cyno IL-13. Anegative control anti-IL-4 mAb showed no neutralization in the samebioassay.

Example 12 Re-Humanisation of Anti-IL-13 mAb Light Chain 12.1Re-Humanisation

The light chain CDRs of the murine antibody 6A1 (The light chain ofwhich is set out in SEQ ID NO:59) were re-grafted onto new frameworks inorder to improve the expression of some anti-IL-13 mAb-anti-IL-4 dAbmolecules (BPC1085). Codon optimised light chain variable regionsequences (summarised in Table 13) were constructed de novo using aPCR-based strategy and overlapping oligonucleotides. PCR primers weredesigned to incorporate the signal sequence (SEQ ID NO: 56) and toinclude HindIII and BsiWl restriction sites designed to frame the V_(L)domain and allow cloning into pTT and Rln mammalian expression vectorscontaining the human kappa C region. Table 13 summarises there-humanised light chains that have been constructed.

TABLE 13 Light SEQ ID NO: SEQ ID NO: chain Back Nucleotide Amino acidname Description Backbone Mutations sequence sequence P0 Anti-IL-13808VL IGKV1 39 + N/A 108 109 kappa light chain JK2 P1 Anti-IL-13 809VLIGKV1 39 + I2V + Q3L 110 111 kappa light chain JK2 Q0 IGKV3 20 + N/A 112113 JK2 Q1 IGKV3 20 + I2V, V3L, L4M, 114 115 JK2 E1D, R45K, I58V

12.2 Molecule Expression in HEK 293 6E Cells

Expression properties of the re-humanised light chains were initiallyexamined in mAb format. Plasmids encoding the A1Y100BVAL1 (SEQ ID NO:54) heavy chain, the existing light chain (SEQ ID NO: 24) and there-humanised light chains were transiently co-transfected into HEK 2936E cells using 293fectin (Invitrogen, 12347019). Plasmids were expressedat small scale (2×0.75 ml culture volumes) to produce antibody. Atryptone feed was added to the cell culture after 24 hours and the cellswere harvested after a further 72 hours. Table 14 summarises all of themAbs which were constructed and expressed.

TABLE 14 Protein SEQ Antibody ID Molecule description ID NO: A1Y100BVAL1H chain: Anti human IL-13 Y100b V mAb 54 L chain: 586 anti-human IL-13mAb 24 BPC3208 H chain: Anti human IL-13 Y100b V mAb 54 L chain: P0re-humanised anti-human IL-13 108 mAb BPC3211 H chain: Anti human IL-13Y100b V mAb 54 L chain: P1 re-humanised anti-human IL-13 110 mAb BPC3219H chain: Anti human IL-13 Y100b V mAb 54 L chain: Q0 re-humanisedanti-human IL-13 112 mAb BPC3220 H chain: Anti human IL-13 Y100b V mAb54 L chain: Q1 re-humanised anti-human IL-13 114 mAb

Antibody expression was assessed directly from the tissue culturesupernatant, by a quantitative immunoassay using a Gyrolab workstation.Antibodies BPC3208 and BPC3211 containing re-humanised light chains(denoted P0 and P1 respectively), exhibited improved expression yieldsin comparison to the A1Y100BVAL1 mAb. Q0 and Q1 light chains (BPC3219and BPC3220) did not improve expression of the anti-IL-13 mAb.Expression data is presented in Table 15.

TABLE 15 Total yield in cell Antibody ID supernatant (μg) A1Y100BVAL119.9 BPC3208 111.0 BPC3211 115.0 BPC3219 16.0 BPC3220 18.312.3 mAb-dAb Expression in HEK 293 6E Cells

As the re-humanised light chains of BPC3208 and BPC3211 exhibitedimproved expression of the anti-IL-13 mAb, they were examined in thecontext of an anti-IL-13 mAb-anti IL-4-dAb. Re-humanised light chains P0and P1 and the 586 (L1) light chain were co-transfected with the829H-GS(TVAAPSGS)₂₋₂₅₆ heavy chain (SEQ ID NO: 96, details summarised inTable 16) into HEK 293 6E cells using 293fectin (Invitrogen, 12347019).Plasmids were expressed at the 50 to 500 ml scale to produce antibodymolecules. A tryptone feed was added to the cell culture after 24 hoursand the cells were harvested after a further 48 hours. Antibodies werepurified using immobilised Protein A columns and quantified by readingabsorbance at 280 nm and where indicated, the purified antibody moleculewas assessed in the assays described in the examples set out below.BPC3214 and BPC3215 were analysed by size exclusion chromatography (SEC)as illustrated in FIGS. 13 and 14.

TABLE 16 Protein DNA Antibody Alternative SEQ ID SEQ ID names LinkerMolecule description NO: ID NO: BPC1085 829H- GS(TVAAPSGS)₂H chain: Anti-human IL-13 96 97 GS(TVAAPSGS)₂- mAb heavy chain Y100B V-256 GS(TVAAPSGS)₂-DOM9- 155-256 L chain: 586 anti-human IL- 24 2513 mAb light chain BPC3214 808H- GS(TVAAPSGS)₂ H chain: Anti-human IL-1396 97 GS(TVAAPSGS)₂- mAb heavy chain Y100B V- 256 GS(TVAAPSGS)₂-DOM9-155-256 L chain: P0 re-humanised 108 109 anti-human IL-13 mAb BPC3215809H- GS(TVAAPSGS)₂ H chain: Anti-human IL-13 96 97 GS(TVAAPSGS)₂-mAb heavy chain Y100B V- 256 GS(TVAAPSGS)₂-DOM9- 155-256L chain: P1 re-humanised 110 111 anti-human IL-13 mAb

Consistent with observations in the mAb format, the mAbdAbs containingthe re-humanised light chains (BPC3214 and BPC3215) exhibited improvedexpression in comparison to BPC1085. Representative expression data issummarised in table 17 5.

TABLE 17 Yield of purified Antibody ID mAbdAb (μg/ml) BPC1085 5.6BPC3214 9.8 BPC3215 7.2

12.4 Human IL-13 Binding ELISA

Purified BPC3214 and BPC3215 were tested for binding to human IL-13 incomparison to BPC1085 (described in Example 10) via a direct bindingELISA. Anti-IL-13 mAb A1Y100BVAL1 and anti-IL-4 mAb were also examinedas positive and negative controls respectively. 96-well high bindingplates were coated with 50 μl/well of recombinant E. coli-expressedhuman IL-13 (Batch number: GRITS31061) at 5 μg/ml and incubated at +4°C. overnight. All subsequent steps were carried out at room temperature.The plates were washed 3 times with phosphate-buffered saline with 0.05%of Tween-20. 100 μL of blocking solution (1% BSA in phosphate-bufferedsaline with 0.05% of Tween-20) was added to each well and the plateswere incubated for at least 1 hour at room temperature. Another washstep was then performed. The purified antibodies were successivelydiluted across the plates in blocking solution. After 1 hour incubation,the plate was washed. Goat anti-human kappa light chain specificperoxidase conjugated antibody was diluted in blocking solution to 0.75μg/ml and 50 μl was added to each well. The plates were incubated forone hour. After another two wash steps, 50 μl of OPD (o-phenylenediaminedihydrochloride) SigmaFast substrate solution was added to each well andthe reaction was stopped by addition of 50 μL of 3M sulphuric acid.Absorbance was read at 490 nm using the VersaMax Microplate Reader(Molecular Devices) using a basic endpoint protocol. These data areshown in FIG. 15. Direct binding ELISA confirmed that BPC3214 andBPC3215 bind to human IL-13. BPC3214 and BPC3215 exhibit similar IL-13binding potency to BPC1085. Positive control anti-IL-13 mAb A1Y100BVAL1also showed binding to recombinant IL-13 whereas negative controlanti-IL-4 mAb demonstrated no binding to human IL-13.

12.5 Human IL-4 Binding ELISA

Purified BPC3214 and BPC3215 were also tested for binding to recombinantE. coli-expressed human IL-4 in a direct binding ELISA. An ELISA wasperformed as described in example 4, coating 96-well high binding plateswith 50 μl/well of recombinant E. coli-expressed human IL-4 at 5 μg/mland incubated at +4° C. overnight. These data are shown in FIG. 16.Direct binding ELISA confirms that BPC3214 and BPC3215 bind to humanIL-4. BPC1085 was also examined. BPC3214 exhibits similar IL-4 bindingpotency to BPC1085. Positive control anti-IL-4 mAb also showed bindingto recombinant IL-4 whereas negative control anti-IL-13 mAb A1Y100BVAL1demonstrated no binding to human IL-4.

Example 13 Binding Affinity of mAbdAbs Comprising the Original IL-13 mAbCDRH3 (BPC2222, BPC2223 & BPC2230-2231) for IL-13 and IL-4 as Assessedby BIAcore™ Analysis Method

Protein A was immobilised on a Cl chip by primary amine coupling; thissurface was used as a capture surface for the antibody molecules to betested. Recombinant E. coli-expressed human IL13 was used at 256, 64,16, 4, and 1 nM, recombinant E. coli-expressed human IL4 was used at 64,16, 4, 1 and 0.25 nM, with 0 nM (i.e. buffer alone) used to doublereference the binding curves of both IL4 and IL13 binding. Regenerationthe Protein A surface was with 100 mM Phosphoric acid. The assay was runat 25° C. using HBS-EP as running buffer. The data was fitted to 1:1model inherent to the Biacore T100 analysis software.

The results of binding to human IL13 are shown in Table 18 and theresult of binding to human IL4 are shown in Table 19.

TABLE 18 Molecule name ka(M/s) kd(1/s) KD (nM) BPC2222 1.31E+06 4.93E−040.376 BPC2223 1.32E+06 4.90E−04 0.372 BPC2230 1.31E+06 4.88E−04 0.373BPC2231 1.30E+06 5.13E−04 0.394

TABLE 19 Molecule name ka(M/s) kd(1/s) KD (nM) BPC2222 1.06E+05 1.09E−041.027 BPC2223 8.59E+06 1.56E−04 0.018 BPC2230* 2.48E+07 2.48E−04 0.010BPC2231* 4.03E+07 2.31E−04 0.006 *The on-rate for BPC2230 and 2231 arebeyond the sensitivity of Biacore, but the fact that we cannotaccurately analyse this data does indicate that the interaction with IL4is likely to be of high affinity with a fast on-rate.

Example 14 Binding Affinity of mAbdAbs Comprising the Original IL-13 mAbCDRH3 (BPC2222, BPC2231) & Variant Anti-IL-13 mAb CDRH3 (BPC1085-1087)for IL-13 and IL-4 as Assessed by BIAcore™ Analysis Method

Protein A was immobilised on a CM5 chip by primary amine coupling; thissurface was used as a capture surface for the antibody molecules to betested. Recombinant E. coli-expressed human IL13 was used at 256 nMonly, Recombinant E. coli-expressed human IL4 was used at 64, 16, 4 and1 nM, with 0 nM (i.e. buffer alone) used to double reference the bindingcurves for both IL4 and IL13 binding. Regeneration the Protein A surfacewas with 50 mM NaOH. The assay was run at 25° C. using HBS-EP as runningbuffer. The data was fitted to 1:1 model inherent to the Biacore T100analysis software.

The results of binding to human IL13 are shown in Table 20, and theresults of binding to human IL4 are shown in Table 21.

TABLE 20 Molecule name ka(M/s) kd(1/s) KD (nM) BPC2222 1.64E+05 5.15E−050.314 BPC2231 5.36E+08 1.16E−03 0.002 BPC1085 1.87E+07 8.97E−04 0.048BPC1086 7.99E+07 1.64E−03 0.021 BPC1087 9.86E+07 1.79E−03 0.018 On-ratefor BPC1086 and BPC1087 are beyond the sensitivity of Biacore, but thefact that we cannot accurately analyse this data does indicate that theinteraction with IL4 is likely to be of high affinity with a faston-rate.

TABLE 21 Molecule name ka(M/s) kd(1/s) KD (nM) BPC2222 1.44E+06 4.44E−040.308 BPC2231 1.56E+06 4.95E−04 0.316 BPC1085 1.20E+06 6.39E−05 0.053BPC1086 1.28E+06 6.57E−05 0.051 BPC1087 1.13E+06 6.42E−05 0.057

Example 15 Binding Affinity of mAbdAbs comprising a number of variantanti-IL-13 mAb CDRH3 (BPC1085, BPC1090-BPC1095, & BPC1108-BPC1119) forIL-4 as assessed by BIAcore™ analysis Method (Human IL-4 BindingAffinity)

Protein A was immobilised on a CM5 chip by primary amine coupling; thissurface was used as a capture surface for the antibody molecules to betested.

Recombinant E. coli-expressed human IL4 was used at 64, 16, 4, 1 and0.25 nM. All binding curves were double referenced with a 0 nM injection(i.e. buffer alone). Regeneration of the Protein A surface was with 50mM NaOH. The assay was run at 25° C. using HBS-EP as running buffer. Thedata was fitted to 1:1 model inherent to the Biacore T100 analysissoftware.

The results of binding to human IL4 are shown in Table 22.

TABLE 22 Molecule name ka(M/s) kd(1/s) KD (nM) BPC1085 1.07E+07 8.34E−040.078 BPC1090 4.34E+06 4.16E−04 0.096 BPC1091 6.05E+06 1.02E−03 0.168BPC1092 2.27E+06 3.89E−03 1.713 BPC1093 3.84E+06 2.11E−04 0.055 BPC10941.46E+06 3.04E−03 2.078 BPC1095 6.78E+06 3.18E−02 4.687 BPC1108 4.38E+061.23E−03 0.281 BPC1109 1.20E+06 1.77E−01 147.300 BPC1110 1.07E+061.73E−03 1.626 BPC1111 2.98E+06 1.11E−04 0.037 BPC1112* 1.14E+083.31E+00 28.980 BPC1113 no binding seen BPC1114 3.89E+06 2.55E−03 0.656BPC1115** 1.44E+08 3.29E−01 2.292 BPC1116 6.82E+06 2.74E−03 0.402BPC1117 3.95E+06 9.13E−04 0.231 BPC1118 6.39E+06 2.58E−03 0.405 BPC1119no binding seen *BPC1112 data has a positive off-rate as a result of themachine being unable to calculate real off-rate, possibly due to thefact it is so rapid, in addition the on-rate for this construct isbeyond what is measurable by Biacore, but construct is a very poorbinder to IL4. *BPC1115 also has an impossible on-rate, it is outsidescope of BIAcore to calculate affinity.

Example 16 IL-13 Binding Affinity of mAbdAbs comprising the re-humanisedlight chain (BPC3214 & BPC3215) compared to the original light chain(BPC1085) as assessed by BIAcore™ analysis Method (Human and Cyno IL-13Binding Affinity)

Protein A was immobilised on a CM5 chip by primary amine coupling; thissurface was used as a capture surface for the antibody molecules to betested. Recombinant E. coli-expressed human IL13 and cyno IL13 were used64, 16, 4, 1 and 0.25 nM. All binding curves were double referenced witha 0 nM injection (i.e. buffer alone).

Regeneration of the Protein A surface was with 50 mM NaOH. The assay wasrun at 25° C. using HBS-EP as running buffer. The data was fitted to 1:1model inherent to the Biacore T100 analysis software.

The results of binding to human and cyno IL13 are shown in Table 23.

TABLE 23 Molecule name ka(M/s) kd(1/s) KD (nM) Comment Binding to humanIL13 BPC3214 7.749E+05 7.18E−05 0.093 BPC3215 8.220E+05 5.23E−05 0.064BPC1085 8.652E+05 6.34E−05 0.073 Binding to cyno IL13 BPC3214* 3.214E+055.07E−06 0.015770 Impossible off-rates BPC3215* 3.283E+05 3.13E−060.009546 Impossible off-rates BPC1085* 3.552E+05 5.19E−10 0.000001Impossible off-rates *The off-rates (ka) for cyno IL13 binding toBPC3214, BPC3215 and BPC1085 are beyond the sensitivity of the BiacoreT100, this indicates the dissociation rate is very slow and that theinteraction is likely to be of very high affinity.

Example 17 Stressor Studies of mAbdAbs with and without the Mutated dAb

A number of mAbdAbs were placed in PBS or 50 mM acetate buffer andincubated at 37° C. for up to 14 days. They were then analysed forpresence of a visual precipitate, soluble aggregate and adherence toconcentration stability.

The results indicate that the mAbdAbs comprising the mutated dAb(BPC2222, 2223, 2230, 2231) behaved similarly to the non-mutated dAb(BPC1085, 1086, 1087) both catagories of mAbdAb appeared to be stable inboth PBS and acetate buffers over the two week incubation period at 37°C., as indicated by no change in the protein concentration in thesolutions. In addition no or very little change was noted for the levelsof aggregates in the solutions and no precipitation was observed.

Example 18 PK Assessment The pharmacokinetics of BPC1085, BPC1086, andBPC1087 were investigated in separate studies following IVadministration to rats. The PK of BPC1085 was also investigated incynomologus monkeys following IV administration.

The PK of all three molecules in rat and BPC1085 in monkey were found tobe consistent with that of a standard mAb.

BRIEF DESCRIPTION OF FIGURES

FIG. 1: A graph showing the capacity of the mAbdAbs comprising the Y100Bvariants to neutralise human IL-13 in a TF-1 cell proliferation assay

FIG. 2: A graph showing the capacity of the mAbdAbs comprising the Y100Bvariants to neutralise human IL-13 in a TF-1 cell proliferation assay

FIG. 3: SEC trace of BPC2222

FIG. 4: SEC trace of BPC2223

FIG. 5: SEC trace of BPC2230

FIG. 6: SEC trace of BPC2231

FIG. 7: SEC trace of BPC1085

FIG. 8: SEC trace of BPC1086

FIG. 9: SEC trace of BPC1087

FIG. 10: A graph showing binding of purified mAbdAbs (BPC1085, BPC1086and BPC1087) to human IL-4 as determined by ELISA. The IL-4 control mAbis labelled as ‘pascolizumab’.

FIG. 11: A graph showing neutralization of human IL-4 by purifiedmAbdAbs (BPC1085, BPC1086 and BPC1087) to human IL-4 in the TF-1 cellbioassay. The IL-4 control mAb is labelled as ‘pascolizumab’.

FIG. 12: A graph showing neutralization of human IL-13 by purifiedmAbdAbs (BPC1085, BPC1086 and BPC1087) to human IL-13 in the TF-1 cellbioassay. The IL-4 control mAb is labelled as ‘pascolizumab’.

FIG. 13: SEC profile for BPC3214.

FIG. 14: SEC profile for BPC3215.

FIG. 15: A graph showing binding of purified mAbdAbs BPC3214, BPC3215,BPC1085 and control mAbs A1Y100BVAL1 and anti-IL-4 mAb to human IL-13 asdetermined by ELISA. The IL-4 control mAb is labelled as ‘pascolizumab’.

FIG. 16: A graph showing binding of purified mAbdAbs BPC3214, BPC3215,BPC1085 and control mAbs A1Y100BVAL1 and anti-IL-4 mAb to human IL-4 asdetermined by ELISA. The IL-4 control mAb is labelled as ‘pascolizumab’.

FIG. 17: A graph showing binding of transiently expressed mAbdAbs torecombinant E. coli-expressed human IL-4 as determined by ELISA. TheIL-4 control mAb is labelled as ‘pascolizumab’.

Sequence Summary (Table 24) Sequence identifier (SEQ ID NO) amino acidPolynucleotide Description sequence sequence Anti IL13 A1, CDRH1 1 —Anti IL13 A1, CDRH2 2 — Anti IL13 A1, CDRH3 3 — CDRH3 alternative 4 —CDRH3 alternative 5 — CDRH3 alternative 6 — CDRH3 alternative 7 — CDRH3alternative 8 — CDRH3 alternative 9 — CDRH3 alternative 10 — CDRH3alternative 11 — CDRH3 alternative 12 — CDRH3 alternative 13 — CDRH3alternative 14 — CDRH3 alternative 15 — CDRH3 alternative 16 — CDRH3alternative 17 — CDRH3 alternative 18 — Anti IL13 L1, CDRL1 19 — AntiIL13 L1, CDRL2 20 — Anti IL13 L1, CDRL3 21 — Anti IL13 A1 (Heavy Chain)22 23 Anti IL13 L1 (Light Chain) 24 25 Anti IL13 humanised variant A1S95 Trp (Heavy 26 27 Chain) Anti IL13 humanised variant A1 I96 Val(Heavy 28 29 Chain) Anti IL13 humanised variant A1 Y97 Phe (Heavy 30 31Chain) Anti IL13 humanised variant A1 D98 Glu (Heavy 32 33 Chain) AntiIL13 humanised variant A1 H100A Ala (Heavy 34 35 Chain) Anti IL13humanised variant A1 H100A Glu (Heavy 36 37 Chain) Anti IL13 humanisedvariant A1 H100A Gln (Heavy 38 39 Chain) Anti IL13 humanised variant A1H100A Arg (Heavy 40 41 Chain) Anti IL13 humanised variant A1 H100A Ser(Heavy 42 43 Chain) Anti IL13 humanised variant A1 H100A Thr (Heavy 4445 Chain) Anti IL13 humanised variant A1 H100A Val (Heavy 46 47 Chain)Anti IL13 humanised variant A1 Y100B Ala (Heavy 48 49 Chain) Anti IL13humanised variant A1 Y100B Ile (Heavy 50 51 Chain) Anti IL13 humanisedvariant A1 Y100B Trp (Heavy 52 53 Chain) Anti IL13 humanised variant A1Y100B Val (Heavy 54 55 Chain) Signal sequence 56 — Human IL13 57 —Murine 6A1 VH 58 — Murine 6A1 VL 59 — Alternative CDRH1 (Chothia andKabat numbering) 60 — Alternative CDRH1 (Chothia and Kabat numbering) 61— A1Y100BAla H-TVAAPS-210 (Heavy chain) 62 63 A1 Y100BIle H-TVAAPS-210(Heavy chain) 64 65 A1Y100BTrp H-TVAAPS-210 (Heavy chain) 66 67A1Y100BVAl H-TVAAPS-210 (Heavy chain) 68 69 A1Y100BAla H-ASTKGPS-210(Heavy chain) 70 71 A1 Y100BIle H-ASTKGPS-210 (Heavy chain) 72 73A1Y100BTrp H-ASTKGPS-210 (Heavy chain) 74 75 A1Y100BVal H-ASTKGPS-210(Heavy chain) 76 77 DOM9-155-25 78 DOM9-155-147 79 DOM9-155-154 80DOM9-112-210 81 Linker 82 Linker 83 Linker 84 Linker 85 Linker 86 Linker87 147-TVAAPS-586 Y100B V Heavy chain 88 147-ASTKG-586 Y100B V Heavychain 89 154-TVAAPS-586 Y100B V Heavy chain 90 154-ASTKG-586 Y100B VHeavy chain 91 Linker 92 Linker 93 DOM9-155-154 L89Q (aka DOM9-155-256)94 95 829H-GS(TVAAPSGS)₂-154 L89Q 96 97 829H-GS(TVAAPSGS)₃-154 L89Q 9899 829H-GS(TVAAPSGS)₄-154 L89Q 100 101 829H-(TVAAPS)₂GS-154 L89Q 102 103829H-(TVAAPS)₃GS-154 L89Q 104 105 829H-(TVAAPS)₄GS-154 L89Q 106 107 P0108 109 P1 110 111 Q0 112 113 Q1 114 115 586H-TVAAPS-154 (H chain) 116Anti-human IL-13 mAb heavy chain Y100B_V- 117 (TVAAPS)₂GS-DOM9-155-154(89G) Anti-human IL-13 mAb heavy chain Y100B_V- 118(TVAAPS)₂GS-DOM9-155-154 (89S) Anti-human IL-13 mAb heavy chain Y100B_V-119 (TVAAPS)₂GS-DOM9-155-154 (89H) Anti-human IL-13 mAb heavy chainY100B_V- 120 (TVAAPS)₂GS-DOM9-155-154 (89M) Anti-human IL-13 mAb heavychain Y100B_V- 121 (TVAAPS)₂GS-DOM9-155-154 (89A) Anti-human IL-13 mAbheavy chain Y100B_V- 122 (TVAAPS)₂GS-DOM9-155-154 (89T) Anti-human IL-13mAb heavy chain Y100B_V- 123 (TVAAPS)₂GS-DOM9-155-154 (89C) Anti-humanIL-13 mAb heavy chain Y100B_V- 124 (TVAAPS)₂GS-DOM9-155-154 (89R)Anti-human IL-13 mAb heavy chain Y100B_V- 125 (TVAAPS)₂GS-DOM9-155-154(89W) Anti-human IL-13 mAb heavy chain Y100B_V- 126(TVAAPS)₂GS-DOM9-155-154 (89E) Anti-human IL-13 mAb heavy chain Y100B_V-127 (TVAAPS)₂GS-DOM9-155-154 (89K) Anti-human IL-13 mAb heavy chainY100B_V- 128 (TVAAPS)₂GS-DOM9-155-154 (89D) Anti-human IL-13 mAb heavychain Y100B_V- 129 (TVAAPS)₂GS-DOM9-155-154 (89N) Anti-human IL-13 mAbheavy chain Y100B_V- 130 (TVAAPS)₂GS-DOM9-155-154 (89Y) Anti-human IL-13mAb heavy chain Y100B_V- 131 (TVAAPS)₂GS-DOM9-155-154 (89V) Anti-humanIL-13 mAb heavy chain Y100B_V- 132 (TVAAPS)₂GS-DOM9-155-154 (89I)Anti-human IL-13 mAb heavy chain Y100B_V- 133 (TVAAPS)₂GS-DOM9-155-154(89F) Anti-human IL-13 mAb heavy chain Y100B_V- 134(TVAAPS)₂GS-DOM9-155-154 (89P) Anti-IL-13 mAb heavy chain-GS(TVAAPSGS)₁-135 — DOM9-155-154 Anti-IL-13 mAb heavy chain-GS(TVAAPSGS)₂- 136 —DOM9-155-154 Anti-IL-13 mAb heavy chain-GS(TVAAPSGS)₃- 137 —DOM9-155-154 Anti-IL-13 mAb heavy chain-GS(TVAAPSGS)₄- 138 DOM9-155-154GS(TVAAPSGS)₁ 139 GS(TVAAPSGS)₂ 140 GS(TVAAPSGS)₃ 141 GS(TVAAPSGS)₄ 142GS(TVAAPSGS)₅ 143 GS(TVAAPSGS)₆ 144 (TVAAPS)₂(GS)₁ 145 (TVAAPS)₃(GS)₁146 Anti-human IL-13 mAb heavy chain Y100B_V- 147(TVAAPS)₂GS-DOM9-155-154 (89M)

SEQUENCES SEQ ID NO: 1 DTYMH SEQ ID NO: 2 TIDPANGNTKYVPKFQG SEQ ID NO: 3SIYDDYHYDDYYAMDY SEQ ID NO: 4 WIYDDYHYDDYYAMDY SEQ ID NO: 5SVYDDYHYDDYYAMDY SEQ ID NO: 6 SIFDDYHYDDYYAMDY SEQ ID NO: 7SIYEDYHYDDYYAMDY SEQ ID NO: 8 SIYDDYAYDDYYAMDY SEQ ID NO: 9SIYDDYEYDDYYAMDY SEQ ID NO: 10 SIYDDYQYDDYYAMDY SEQ ID NO: 11SIYDDYRYDDYYAMDY SEQ ID NO: 12 SIYDDYSYDDYYAMDY SEQ ID NO: 13SIYDDYTYDDYYAMDY SEQ ID NO: 14 SIYDDYVYDDYYAMDY SEQ ID NO: 15SIYDDYHADDYYAMDY SEQ ID NO: 16 SIYDDYHIDDYYAMDY SEQ ID NO: 17SIYDDYHWDDYYAMDY SEQ ID NO: 18 SIYDDYHVDDYYAMDY SEQ ID NO: 19RSSQNIVHINGNTYLE SEQ ID NO: 20 KISDRFS SEQ ID NO: 21 FQGSHVPWT SEQ IDNO: 22 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 23 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 24 DIVMTQSPLSLPVTPGEPASISCRSSQNIVHINGNTYLEWYLQKPGQSPRLLIYKISDRFSGVPDRFSGSGSGTDFTLKISRVEADDVGIYYCFQGSHVPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 25GACATCGTGATGACCCAGTCTCCTCTGAGCCTCCCCGTGACCCCCGGCGAACCAGCCAGCATCTCCTGCAGAAGCAGCCAGAACATCGTGCACATCAACGGCAACACCTACCTGGAGTGGTACCTGCAAAAGCCCGGCCAGAGCCCCAGGCTGCTGATCTACAAGATCAGCGACAGGTTCAGCGGCGTGCCCGATAGGTTCAGCGGCAGCGGCAGCGGCACCGACTTCACCCTGAAGATCAGCAGGGTGGAGGCCGACGACGTGGGCATCTACTACTGCTTCCAGGGCAGCCACGTCCCCTGGACTTTCGGACAGGGCACCAAGCTGGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGC SEQ ID NO: 26QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARWIYDDYHYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 27 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGTGGATCTACGACGACTACCACTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 28 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSVYDDYHYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 29 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCGTCTACGACGACTACCACTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 30 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIFDDYHYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 31 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTTTGACGACTACCACTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 32 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYEDYHYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 33 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGAGGACTACCACTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 34 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYAYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 35 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACGCGTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 36 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYEYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 37 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACGAGTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 38 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYQYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 39 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCAGTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 40 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYRYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 41 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACAGGTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 42 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYSYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 43 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACTCCTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 44 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYTYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 45 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACACGTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 46 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYVYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 47 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACGTGTACGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 48 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHADDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 49 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGCGGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 50 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHIDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 51 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACATTGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 52 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHWDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 53 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACTGGGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 54 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ IDNO: 55 CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGTCGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAG SEQ IDNO: 56 MGWSCIILFLVATATGVHS SEQ ID NO: 57GPVPPSTALRELIEELVNITQNQKAPLCNGSMVWSINLTAGMYCAALESLINVSGCSAIEKTQRMLSGFCPHKVSAGQFSSLHVRDTKIEVAQFVKDLLLHLKKLFREGRFN SEQ ID NO: 58EIQLQQSVAELVRPGASVRLSCTASGFYIKDTYMHWVIQRPEQGLEWIGTIDPANGNTKYVPKFQGKATITADTSSNTAYLRLSSLTSEDTAIYYCARSIYDDYHYDDYYAMDYWGQGTSVTVSS SEQ ID NO:59 DVLMTQTPLSLPVSLGDQASISCRSSQNIVHINGNTYLEWYLQKPGQSPKLLIYKISDRFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPWTFGGGTKLEIK SEQ ID NO: 60 FYIKDTYMHSEQ ID NO: 61 GFYIKDTYMH SEQ ID 62: A1Y100BAla H-TVAAPS-210 (Proteinsequence)QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHADDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSEVQLLESGGGLVQPGGSLRLSCAASGFTFRNFGMGWVRQAPGKGLEWVSWIISSGTETYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSLGRFDYWGQGTLVTVSS SEQ ID 63:A1Y100BAla H-TVAAPS-210 (DNA sequence)CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGCGGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCTAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGACCGTGGCCGCCCCCTCGGAAGTGCAGCTCCTGGAGAGCGGCGGCGGCCTGGTGCAGCCCGGCGGCAGCCTGAGGCTGAGCTGCGCCGCTAGCGGCTTCACCTTCAGGAACTTCGGCATGGGCTGGGTCAGGCAGGCCCCCGGCAAGGGCCTGGAGTGGGTCAGCTGGATCATCAGCTCCGGCACCGAGACCTACTACGCCGACAGCGTGAAGGGCAGGTTCACCATCAGCCGCGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGGGCCGAGGACACCGCCGTCTACTACTGCGCCAAGAGCCTGGGCAGGTTCGACTACTGGGGACAGGGGACCCTGGTGACTGTGAGCAGC SEQ ID 64: A1Y100BIleH-TVAAPS-210 (Protein sequence)QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHIDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSEVQLLESGGGLVQPGGSLRLSCAASGFTFRNFGMGWVRQAPGKGLEWVSWIISSGTETYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSLGRFDYWGQGTLVTVSS SEQ ID 65:A1Y100BIle H-TVAAPS-210 (DNA sequence)CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACATTGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCTAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGACCGTGGCCGCCCCCTCGGAAGTGCAGCTCCTGGAGAGCGGCGGCGGCCTGGTGCAGCCCGGCGGCAGCCTGAGGCTGAGCTGCGCCGCTAGCGGCTTCACCTTCAGGAACTTCGGCATGGGCTGGGTCAGGCAGGCCCCCGGCAAGGGCCTGGAGTGGGTCAGCTGGATCATCAGCTCCGGCACCGAGACCTACTACGCCGACAGCGTGAAGGGCAGGTTCACCATCAGCCGCGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGGGCCGAGGACACCGCCGTCTACTACTGCGCCAAGAGCCTGGGCAGGTTCGACTACTGGGGACAGGGGACCCTGGTGACTGTGAGCAGC SEQ ID 66: A1Y100BTrpH-TVAAPS-210 (Protein sequence)QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHWDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSEVQLLESGGGLVQPGGSLRLSCAASGFTFRNFGMGWVRQAPGKGLEWVSWIISSGTETYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSLGRFDYWGQGTLVTVSS SEQ ID 67:A1Y100BTrp H-TVAAPS-210 (DNA sequence)CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACTGGGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCTAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGACCGTGGCCGCCCCCTCGGAAGTGCAGCTCCTGGAGAGCGGCGGCGGCCTGGTGCAGCCCGGCGGCAGCCTGAGGCTGAGCTGCGCCGCTAGCGGCTTCACCTTCAGGAACTTCGGCATGGGCTGGGTCAGGCAGGCCCCCGGCAAGGGCCTGGAGTGGGTCAGCTGGATCATCAGCTCCGGCACCGAGACCTACTACGCCGACAGCGTGAAGGGCAGGTTCACCATCAGCCGCGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGGGCCGAGGACACCGCCGTCTACTACTGCGCCAAGAGCCTGGGCAGGTTCGACTACTGGGGACAGGGGACCCTGGTGACTGTGAGCAGC SEQ ID 68: A1Y100BValH-TVAAPS-210 (Protein sequence)QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSEVQLLESGGGLVQPGGSLRLSCAASGFTFRNFGMGWVRQAPGKGLEWVSWIISSGTETYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSLGRFDYWGQGTLVTVSS SEQ ID 69:A1Y100BVal H-TVAAPS-210 (DNA sequence)CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGTCGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGACCGTGGCCGCCCCCTCGGAAGTGCAGCTCCTGGAGAGCGGCGGCGGCCTGGTGCAGCCCGGCGGCAGCCTGAGGCTGAGCTGCGCCGCTAGCGGCTTCACCTTCAGGAACTTCGGCATGGGCTGGGTCAGGCAGGCCCCCGGCAAGGGCCTGGAGTGGGTCAGCTGGATCATCAGCTCCGGCACCGAGACCTACTACGCCGACAGCGTGAAGGGCAGGTTCACCATCAGCCGCGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGGGCCGAGGACACCGCCGTCTACTACTGCGCCAAGAGCCTGGGCAGGTTCGACTACTGGGGACAGGGGACCCTGGTGACTGTGAGCAGC SEQ ID 70: A1Y100BAlaH-ASTKGPS-210 (Protein sequence)QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHADDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKASTKGPSEVQLLESGGGLVQPGGSLRLSCAASGFTFRNFGMGWVRQAPGKGLEWVSWIISSGTETYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSLGRFDYWGQGTLVTVSS SEQ ID 71:A1Y100BAla H-ASTKGPS-210 (DNA sequence)CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGCGGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGGCCAGCACCAAGGGCCCCTCGGAAGTGCAGCTCCTGGAGAGCGGCGGCGGCCTGGTGCAGCCCGGCGGCAGCCTGAGGCTGAGCTGCGCCGCTAGCGGCTTCACCTTCAGGAACTTCGGCATGGGCTGGGTCAGGCAGGCCCCCGGCAAGGGCCTGGAGTGGGTCAGCTGGATCATCAGCTCCGGCACCGAGACCTACTACGCCGACAGCGTGAAGGGCAGGTTCACCATCAGCCGCGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGGGCCGAGGACACCGCCGTCTACTACTGCGCCAAGAGCCTGGGCAGGTTCGACTACTGGGGACAGGGGACCCTGGTGACTGTGAGCAGC SEQ ID 72: A1Y100BIleH-ASTKGPS-210 (Protein sequence)QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHIDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKASTKGPSEVQLLESGGGLVQPGGSLRLSCAASGFTFRNFGMGWVRQAPGKGLEWVSWIISSGTETYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSLGRFDYWGQGTLVTVSS SEQ ID 73:A1Y100BIle H-ASTKGPS-210 (DNA sequence)CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACATTGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGGCCAGCACCAAGGGCCCCTCGGAAGTGCAGCTCCTGGAGAGCGGCGGCGGCCTGGTGCAGCCCGGCGGCAGCCTGAGGCTGAGCTGCGCCGCTAGCGGCTTCACCTTCAGGAACTTCGGCATGGGCTGGGTCAGGCAGGCCCCCGGCAAGGGCCTGGAGTGGGTCAGCTGGATCATCAGCTCCGGCACCGAGACCTACTACGCCGACAGCGTGAAGGGCAGGTTCACCATCAGCCGCGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGGGCCGAGGACACCGCCGTCTACTACTGCGCCAAGAGCCTGGGCAGGTTCGACTACTGGGGACAGGGGACCCTGGTGACTGTGAGCAGC SEQ ID 74: A1Y100BTrpH-ASTKGPS-210 (Protein sequence)QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHWDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKASTKGPSEVQLLESGGGLVQPGGSLRLSCAASGFTFRNFGMGWVRQAPGKGLEWVSWIISSGTETYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSLGRFDYWGQGTLVTVSS SEQ ID 75:A1Y100BTrpH-ASTKGPS-210 (DNA sequence)CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACTGGGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGGCCAGCACCAAGGGCCCCTCGGAAGTGCAGCTCCTGGAGAGCGGCGGCGGCCTGGTGCAGCCCGGCGGCAGCCTGAGGCTGAGCTGCGCCGCTAGCGGCTTCACCTTCAGGAACTTCGGCATGGGCTGGGTCAGGCAGGCCCCCGGCAAGGGCCTGGAGTGGGTCAGCTGGATCATCAGCTCCGGCACCGAGACCTACTACGCCGACAGCGTGAAGGGCAGGTTCACCATCAGCCGCGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGGGCCGAGGACACCGCCGTCTACTACTGCGCCAAGAGCCTGGGCAGGTTCGACTACTGGGGACAGGGGACCCTGGTGACTGTGAGCAGC SEQ ID 76: A1Y100BValH-ASTKGPS-210 (Protein sequence)QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKASTKGPSEVQLLESGGGLVQPGGSLRLSCAASGFTFRNFGMGWVRQAPGKGLEWVSWIISSGTETYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSLGRFDYWGQGTLVTVSS SEQ ID 77:A1Y100BVal H-ASTKGPS-210 (DNA sequence)CAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGTCGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGGCCAGCACCAAGGGCCCCTCGGAAGTGCAGCTCCTGGAGAGCGGCGGCGGCCTGGTGCAGCCCGGCGGCAGCCTGAGGCTGAGCTGCGCCGCTAGCGGCTTCACCTTCAGGAACTTCGGCATGGGCTGGGTCAGGCAGGCCCCCGGCAAGGGCCTGGAGTGGGTCAGCTGGATCATCAGCTCCGGCACCGAGACCTACTACGCCGACAGCGTGAAGGGCAGGTTCACCATCAGCCGCGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGGGCCGAGGACACCGCCGTCTACTACTGCGCCAAGAGCCTGGGCAGGTTCGACTACTGGGGACAGGGGACCCTGGTGACTGTGAGCAGC SEQ ID NO: 78 = DOM9-155-25DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASTLDSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKR SEQ ID NO: 79 = DOM9-155-147DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLYEGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKR SEQ ID NO: 80 = DOM9-155-154DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKR SEQ ID NO: 81 = DOM9-112-210EVQLLESGGGLVQPGGSLRLSCAASGFTFRNFGMGWVRQAPGKGLEWVSWIISSGTETYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKSLGRFDYWGQGTLVTVSS SEQ ID NO: 82 (G4Slinker) GGGGS SEQ ID NO: 83 (linker) TVAAPS SEQ ID NO: 84 (linker)ASTKGPT SEQ ID NO: 85 (linker) ASTKGPS SEQ ID NO: 86 (linker) GS SEQ IDNO: 87 (linker) TVAAPSGS SEQ ID NO: 88 = 147-TVAAPS-586 Y100B V (Heavychain) DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLYEGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKRGSTVAAPSQVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 89= 147-ASTKG-586 Y100B V (Heavy chain)DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLYEGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKRGSASTKGPSQVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 90= 154-TVAAPS-586 Y100B V (Heavy chain)DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKRGSTVAAPSQVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 91= 154-ASTKG-586 Y100B V (Heavy chain)DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKRGSASTKGPSQVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 92(linker) GSTVAAPS SEQ ID NO: 93 (linker) GSTVAAPSGS SEQ ID NO: 94DOM9-155-154 L89Q (aka DOM9-155-256)DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQEGWGPPTFGQGTKVEIKR SEQ ID NO: 95DOM9-155-154 L89Q (aka DOM9-155-256)GACATCCAGATGACCCAGAGCCCCAGCAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACCATTACCTGCAGGGCCAGCAGGCCCATCAGCGACTGGCTGCACTGGTACCAACAGAAGCCCGGCAAGGCTCCCAAGCTGCTGATCGCCTGGGCCAGCAGCCTGCAGGGAGGCGTGCCCAGCAGGTTTAGCGGCAGCGGCAGCGGCACCGACTTCACCCTCACCATCTCTTCCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGGAGGGCTGGGGGCCCCCTACTTTCGGCCAGGGCACCAAGGTGGAGATCAAGAGG SEQ ID NO: 96 829H-GS(TVAAPSGS)₂-154 L89QQVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSTVAAPSGSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQEGWGPPTFGQGTKVEIKR SEQ ID NO: 97829H-GS(TVAAPSGS)₂-154 L89QCAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGTCGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGGGATCTACCGTGGCAGCACCATCCGGATCTACCGTAGCAGCACCATCCGGATCCGACATCCAGATGACCCAGAGCCCCAGCAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACCATTACCTGCAGGGCCAGCAGGCCCATCAGCGACTGGCTGCACTGGTACCAACAGAAGCCCGGCAAGGCTCCCAAGCTGCTGATCGCCTGGGCCAGCAGCCTGCAGGGAGGCGTGCCCAGCAGGTTTAGCGGCAGCGGCAGCGGCACCGACTTCACCCTCACCATCTCTTCCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGGAGGGCTGGGGGCCCCCTACTTTCGGCCAGGGCACCAAGGTGGAGATCAAG AGG SEQ IDNO: 98 829H-GS(TVAAPSGS)₃-154 L89QQVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSTVAAPSGSTVAAPSGSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQEGWGPPTFGQGTKVEIKR SEQ ID NO: 99829H-GS(TVAAPSGS)₃-154 L89QCAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGTCGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGGGATCTACCGTGGCAGCACCATCAGGATCTACCGTGGCAGCACCATCAGGTTCAACAGTAGCTGCTCCTTCTGGATCCGACATCCAGATGACCCAGAGCCCCAGCAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACCATTACCTGCAGGGCCAGCAGGCCCATCAGCGACTGGCTGCACTGGTACCAACAGAAGCCCGGCAAGGCTCCCAAGCTGCTGATCGCCTGGGCCAGCAGCCTGCAGGGAGGCGTGCCCAGCAGGTTTAGCGGCAGCGGCAGCGGCACCGACTTCACCCTCACCATCTCTTCCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGGAGGGCTGGGGGCCCCCTACTTTCGGCCAGGGCACCAAGGTGGAGATCAAGAGG SEQ ID NO: 100 829H-GS(TVAAPSGS)₄-154 L89QQVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSTVAAPSGSTVAAPSGSTVAAPSGSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQEGWGPPTFGQGTKVEIKR SEQ ID NO: 101829H-GS(TVAAPSGS)₄-154 L89QCAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGTCGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGGGATCTACCGTGGCAGCACCATCAGGATCTACCGTGGCAGCACCATCAGGTTCAACAGTAGCTGCTCCTTCTGGTTCAACAGTAGCTGCTCCTTCTGGATCCGACATCCAGATGACCCAGAGCCCCAGCAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACCATTACCTGCAGGGCCAGCAGGCCCATCAGCGACTGGCTGCACTGGTACCAACAGAAGCCCGGCAAGGCTCCCAAGCTGCTGATCGCCTGGGCCAGCAGCCTGCAGGGAGGCGTGCCCAGCAGGTTTAGCGGCAGCGGCAGCGGCACCGACTTCACCCTCACCATCTCTTCCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGGAGGGCTGGGGGCCCCCTACTTTCGGCCAGGGCACCAAGGTGGAGATCAAGAGG SEQ ID NO: 102829H-(TVAAPS)₂GS-154 L89QQVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQEGWGPPTFGQGTKVEIKR SEQ ID NO: 103829H-(TVAAPS)₂GS-154 L89QCAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGTCGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGACCGTGGCAGCACCATCCACCGTAGCAGCACCATCCGGATCCGACATCCAGATGACCCAGAGCCCCAGCAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACCATTACCTGCAGGGCCAGCAGGCCCATCAGCGACTGGCTGCACTGGTACCAACAGAAGCCCGGCAAGGCTCCCAAGCTGCTGATCGCCTGGGCCAGCAGCCTGCAGGGAGGCGTGCCCAGCAGGTTTAGCGGCAGCGGCAGCGGCACCGACTTCACCCTCACCATCTCTTCCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGGAGGGCTGGGGGCCCCCTACTTTCGGCCAGGGCACCAAGGTGGAGATCAAGAGG SEQ ID NO: 104829H-(TVAAPS)₃GS-154 L89QQVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQEGWGPPTFGQGTKVEIKR SEQ ID NO: 105829H-(TVAAPS)₃GS-154 L89QCAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGTCGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGACCGTGGCAGCACCATCAACCGTGGCAGCACCATCAACAGTAGCTGCTCCTTCTGGATCCGACATCCAGATGACCCAGAGCCCCAGCAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACCATTACCTGCAGGGCCAGCAGGCCCATCAGCGACTGGCTGCACTGGTACCAACAGAAGCCCGGCAAGGCTCCCAAGCTGCTGATCGCCTGGGCCAGCAGCCTGCAGGGAGGCGTGCCCAGCAGGTTTAGCGGCAGCGGCAGCGGCACCGACTTCACCCTCACCATCTCTTCCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGGAGGGCTGGGGGCCCCCTACTTTCGGCCAGGGCACCAAGGTGGAG ATCAAGAGGSEQ ID NO: 106 829H-(TVAAPS)₄GS-154 L89QQVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQEGWGPPTFGQGTKVEIKR SEQ ID NO: 107829H-(TVAAPS)₄GS-154 L89QCAGGTGCAGCTCGTGCAGAGCGGCGCCGAAGTGAAAAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCTCCGGCTTCTACATCAAGGACACCTACATGCACTGGGTCAGGCAGGCTCCTGGCCAGGGCCTGGAGTGGATGGGCACTATCGACCCCGCCAACGGCAACACCAAGTACGTGCCCAAGTTCCAGGGCAGGGTGACCATCACCGCCGATGAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGAGGTCTGAGGACACCGCCGTGTACTATTGCGCCAGGAGCATCTACGACGACTACCACGTCGACGACTACTACGCCATGGACTACTGGGGACAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCAAGACCGTGGCAGCACCATCAACCGTGGCAGCACCATCAACAGTAGCTGCTCCTTCTACAGTAGCTGCTCCTTCTGGATCCGACATCCAGATGACCCAGAGCCCCAGCAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACCATTACCTGCAGGGCCAGCAGGCCCATCAGCGACTGGCTGCACTGGTACCAACAGAAGCCCGGCAAGGCTCCCAAGCTGCTGATCGCCTGGGCCAGCAGCCTGCAGGGAGGCGTGCCCAGCAGGTTTAGCGGCAGCGGCAGCGGCACCGACTTCACCCTCACCATCTCTTCCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCCAGCAGGAGGGCTGGGGGCCCCCTACTTTCGGCCAGGGCACCAAGGTGGAGATCAAGAGG SEQ ID NO: 108DIQMTQSPSSLSASVGDRVTITCRSSQNIVHINGNTYLEWYQQKPGKAPKLLIYKISDRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQGSHVPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 109ATGGGCTGGTCCTGCATCATCCTGTTTCTGGTGGCCACCGCCACCGGCGTGCACAGCGACATCCAGATGACCCAGAGCCCCAGCAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACTATCACCTGCAGGAGCAGCCAGAACATCGTGCACATCAACGGCAACACCTACCTCGAGTGGTACCAGCAGAAACCCGGGAAGGCCCCCAAGCTGCTGATCTACAAGATCAGCGACAGGTTCAGCGGCGTGCCCAGCAGGTTTAGCGGCTCCGGCTCAGGCACCGATTTCACCCTGACCATTAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCTTCCAGGGCTCTCACGTCCCCTGGACCTTCGGCCAGGGCACCAAGCTGGAGATCAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGC SEQ IDNO: 110DVLMTQSPSSLSASVGDRVTITCRSSQNIVHINGNTYLEWYQQKPGKAPKLLIYKISDRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQGSHVPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 111ATGGGCTGGTCCTGCATCATCCTGTTTCTGGTGGCCACCGCCACCGGCGTGCACAGCGACGTGCTGATGACCCAGAGCCCCAGCAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACTATCACCTGCAGGAGCAGCCAGAACATCGTGCACATCAACGGCAACACCTACCTCGAGTGGTACCAGCAGAAACCCGGGAAGGCCCCCAAGCTGCTGATCTACAAGATCAGCGACAGGTTCAGCGGCGTGCCCAGCAGGTTTAGCGGCTCCGGCTCAGGCACCGATTTCACCCTGACCATTAGCAGCCTGCAGCCCGAGGACTTCGCCACCTACTACTGCTTCCAGGGCTCTCACGTCCCCTGGACCTTCGGCCAGGGCACCAAGCTGGAGATCAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGC SEQ IDNO: 112EIVLTQSPGTLSLSPGERATLSCRSSQNIVHINGNTYLEWYQQKPGQAPRLLIYKISDRFSGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCFQGSHVPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 113ATGGGCTGGTCCTGCATCATCCTGTTTCTGGTGGCCACCGCCACCGGCGTGCACAGCGAGATCGTGCTGACCCAGAGCCCTGGCACACTGAGCCTGAGCCCCGGAGAGAGGGCCACCCTGAGCTGCAGGTCTAGCCAGAACATCGTGCACATCAACGGCAACACCTACCTGGAGTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGGCTGCTGATCTACAAGATCAGCGACAGGTTCAGCGGCATCCCCGACAGGTTTAGCGGCAGCGGCAGCGGCACCGACTTCACCCTGACCATTAGCAGGCTGGAGCCCGAGGACTTCGCCGTGTACTACTGCTTCCAGGGGAGCCACGTGCCCTGGACCTTCGGCCAGGGCACCAAGCTCGAAATCAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGC SEQ IDNO: 114DVLMTQSPGTLSLSPGERATLSCRSSQNIVHINGNTYLEWYQQKPGQAPKLLIYKISDRFSGVPDRFSGSGSGTDFTLTISRLEPEDFAVYYCFQGSHVPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 115GACGTGCTGATGACCCAGAGCCCTGGCACACTGAGCCTGAGCCCCGGAGAGAGGGCCACCCTGAGCTGCAGGTCTAGCCAGAACATCGTGCACATCAACGGCAACACCTACCTGGAGTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAAGCTGCTGATCTACAAGATCAGCGACAGGTTCAGCGGCGTGCCCGACAGGTTTAGCGGCAGCGGCAGCGGCACCGACTTCACCCTGACCATTAGCAGGCTGGAGCCCGAGGACTTCGCCGTGTACTACTGCTTCCAGGGGAGCCACGTGCCCTGGACCTTCGGCCAGGGCACCAAGCTCGAAATCAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGC SEQ ID NO: 116 = 586H-TVAAPS-154 (H chain)QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKR SEQ ID NO: 117QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCGQEGWGPPTFGQGTKVEIKR SEQ ID NO: 118QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCSQEGWGPPTFGQGTKVEIKR SEQ ID NO: 119QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQEGWGPPTFGQGTKVEIKR SEQ ID NO: 120QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCMQEGWGPPTFGQGTKVEIKR SEQ ID NO: 121QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCAQEGWGPPTFGQGTKVEIKR SEQ ID NO: 122QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCTQEGWGPPTFGQGTKVEIKR SEQ ID NO: 123QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCCQEGWGPPTFGQGTKVEIKR SEQ ID NO: 124QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCRQEGWGPPTFGQGTKVEIKR SEQ ID NO: 125QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCWQEGWGPPTFGQGTKVEIKR SEQ ID NO: 126QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCEQEGWGPPTFGQGTKVEIKR SEQ ID NO: 127QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCKQEGWGPPTFGQGTKVEIKR SEQ ID NO: 128QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCDQEGWGPPTFGQGTKVEIKR SEQ ID NO: 129QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCNQEGWGPPTFGQGTKVEIKR SEQ ID NO: 130QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCYQEGWGPPTFGQGTKVEIKR SEQ ID NO: 131QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCVQEGWGPPTFGQGTKVEIKR SEQ ID NO: 132QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCIQEGWGPPTFGQGTKVEIKR SEQ ID NO: 133QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQEGWGPPTFGQGTKVEIKR SEQ ID NO: 134QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHVDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCPQEGWGPPTFGQGTKVEIKR SEQ ID NO: 135QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSTVAAPSGS DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKR SEQ ID NO: 136QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSTVAAPSGSTVAAPSGS DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKR SEQ ID NO:137 QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSTVAAPSGSTVAAPSGSTVAAPSGS DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKV EIKRSEQ ID NO: 138QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARSIYDDYHYDDYYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGSTVAAPSGSTVAAPSGSTVAAPSGSTVAAPSGS DIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQEGWGPPTFGQGTKVEIKR SEQ ID NO: 139 GSTVAAPSGS SEQ ID NO: 140 GSTVAAPSGSTVAAPSGSSEQ ID NO: 141 GSTVAAPSGSTVAAPSGSTVAAPSGS SEQ ID NO: 142GSTVAAPSGSTVAAPSGSTVAAPSGSTVAAPSGS SEQ ID NO: 143GSTVAAPSGSTVAAPSGSTVAAPSGSTVAAPSGSTVAAPSGS SEQ ID NO: 144GSTVAAPSGSTVAAPSGSTVAAPSGSTVAAPSGSTVAAPSGSTVAAPSGS SEQ ID NO: 145TVAAPSTVAAPSGS SEQ ID NO: 146 TVAAPSTVAAPSTVAAPSGS SEQ ID NO: 147QVQLVQSGAEVKKPGSSVKVSCKASGFYIKDTYMHWVRQAPGQGLEWMGTIDPANGNTKYVPKFQGRVTITADESTSTAYMELSSLRSEDSAVYYCARSIYDDYHVDDYYAMDYLGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKTVAAPSTVAAPSGSDIQMTQSPSSLSASVGDRVTITCRASRPISDWLHWYQQKPGKAPKLLIAWASSLQGGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCMQEGWGPPTFGQGTKVEIKR

1-31. (canceled)
 32. An antigen binding protein which binds human IL-13and which comprises the CDRH3 SIYDDYHYDDYYAMDY(SEQ ID NO: 3), whereinCDRH3 comprises one or more of the following substitutions: a) S95 inposition 1 is substituted for tryptophan (W) b) I96 in position 2 issubstituted for valine (V) c) Y97 in position 3 is substituted forphenylalanine (F) d) D98 in position 4 is substituted for glutamine (E)e) H100A in position 7 is substituted for alanine (A), glutamic acid(E), glutamine (Q), Arginine (R), Serine (S), threonine (T) or valine(V), and f) Y100B in position 8 is substituted for alanine (A),isoleucine, (I), tryptophan (W) or valine (V).
 33. The antigen bindingprotein according to claim 32 wherein CDRH3 is substituted at Y100B inposition 8 to an amino acid selected from alanine (A), isoleucine, (I),tryptophan (W) or valine (V).
 34. The antigen binding protein accordingto claim 32 which binds human IL-13 and which comprises a CDRH3 sequenceselected from those set out in SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6,SEQ ID NO: 7, SEQ ID NO:8 and SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO:11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14 and SEQ ID NO: 15, SEQ IDNO: 16, SEQ ID NO: 17 and SEQ ID NO:
 18. 35. The antigen binding proteinaccording to claim 32 wherein said antigen binding protein furthercomprises the following CDR sequences: CDRH1: SEQ ID NO:1, CDRH2: SEQ IDNO: 2; CDRL1: SEQ ID NO:19; CDRL2: SEQ ID NO:20; and CDRL3: SEQ IDNO:21.
 36. The antigen binding protein according to claim 32 whereinsaid antigen binding protein comprises the following CDRs: CDRH1: SEQ IDNO:1, CDRH2: SEQ ID NO: 2; CDRH3: SEQ ID NO: 18 CDRL1: SEQ ID NO:19;CDRL2: SEQ ID NO:20; and CDRL3: SEQ ID NO:21.
 37. The antigen bindingprotein according to claim 32 wherein the antigen binding proteincomprises a humanised antibody of IgG isotype.
 38. The antigen bindingprotein according to claim 32 comprising a heavy chain selected from SEQID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34,SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO:44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52 and SEQID NO: 54; and the light chain of SEQ ID NO: 24, 108, 110, 112 and 114.39. The antigen binding protein according to claim 38 comprising theheavy chain of SEQ ID NO: 54 and the light chain of SEQ ID NO:108. 40.The antigen binding protein according to claim 32 which is capable ofbinding to IL-13 and which is also capable of binding to at least one ofIL-4 and IL-5.
 41. The antigen binding protein according to claim 40which comprises at least one dAb selected from SEQ ID NO: 78, 79, 80, 81and
 94. 42. The antigen binding protein according to claim 41 whichcomprises a heavy chain selected from any one of SEQ ID NO: 62, 64, 66,68, 70, 72, 74, 76, 96, 98, 100, 102, 104, 106, and 117-138 and thelight of SEQ ID NO: 24, 108, 110, 112 and
 114. 43. The antigen bindingprotein according to claim 42 which comprises a heavy chain selectedfrom any one of SEQ ID NO: 96, 98, 100, 102, 104, 106 and the light ofSEQ ID NO: 24, 108 and
 110. 44. A recombinant transformed or transfectedhost cell comprising a first and second vector, said first vectorcomprising a polynucleotide encoding a heavy chain of an antibodyaccording to any preceding claim and said second vector comprising apolynucleotide encoding a light chain of claim
 32. 45. A pharmaceuticalcomposition comprising an antigen binding protein of claim 32 and apharmaceutically acceptable carrier.
 46. A method of treating a humanpatient with alergic asthma, severe asthma, difficult asthma, brittleasthma, nocturnal asthma, premenstrual asthma, steroid resistant asthma,steroid dependent asthma, aspirin induced asthma, adult-onset asthma,paediatric asthma, atopic dermatitis, allergic rhinitis, Crohn'sdisease, COPD, fibrotic diseases or disorders such as idiopathicpulmonary fibrosis, progressive systemic sclerosis, hepatic fibrosis,hepatic granulomas, schistosomiasis, leishmaniasis, diseases of cellcycle regulation such as Hodgkins disease, B cell chronic lymphocyticleukaemia which method comprises the step of administering atherapeutically effective amount of an antigen binding protein of 32.47. Use of an antigen binding protein according to claim 32 in thepreparation of a medicament for treatment or prophylaxis of Allergicasthma, severe asthma, difficult asthma, brittle asthma, nocturnalasthma, premenstrual asthma, steroid resistant asthma, steroid dependentasthma, aspirin induced asthma, adult-onset asthma, paediatric asthma,atopic dermatitis, allergic rhinitis, Crohn's disease, COPD, fibroticdiseases or disorders such as idiopathic pulmonary fibrosis, progressivesystemic sclerosis, hepatic fibrosis, hepatic granulomas,schistosomiasis, leishmaniasis, diseases of cell cycle regulation suchas Hodgkins disease, B cell chronic lymphocytic leukaemia.
 48. Theantigen binding protein according to claim 32 for use in the treatmentor prophylaxis of Allergic asthma, severe asthma, difficult asthma,brittle asthma, nocturnal asthma, premenstrual asthma, steroid resistantasthma, steroid dependent asthma, aspirin induced asthma, adult-onsetasthma, paediatric asthma, atopic dermatitis, allergic rhinitis, Crohn'sdisease, COPD, fibrotic diseases or disorders such as idiopathicpulmonary fibrosis, progressive systemic sclerosis, hepatic fibrosis,hepatic granulomas, schistosomiasis, leishmaniasis, diseases of cellcycle regulation such as Hodgkins disease, B cell chronic lymphocyticleukaemia.